S1
Microbe-focused glycan array screening platform
Andreas Geissner, Anika Reinhardt, Christoph Rademacher, Timo Johannssen,
João Monteiro, Bernd Lepenies, Michel Thépaut, Franck Fieschi, Jana Mrázková,
Michaela Wimmerova, Frank Schuhmacher, Sebastian Götze, Dan Grünstein,
Xiaoqiang Guo, Heung Sik Hahm, Jeyakumar Kandasamy, Daniele Leonori,
Christopher E. Martin, Sharavathi G. Parameswarappa, Sandip Pasari, Mark K. Schlegel,
Hidenori Tanaka, Guozhi Xiao, You Yang, Claney L. Pereira, Chakkumkal Anish,
and Peter H. Seeberger
SI Appendix
www.pnas.org/cgi/doi/10.1073/pnas.1800853116
S2
1. Experimental Procedures
Microarray printing
Glycans were dissolved at a final concentration of 100 µM in sterile printing buffer (50 mM
sodium phosphate, pH 8.5). For some rare exceptions of glycans containing base labile groups
(indicated in SI Appendix Table S2), printing solutions were prepared in phosphate buffered
saline (PBS, pH 7.4). Approximately 400 pL of the substances per spot were printed in 16 field
format (overall 284 spots per field) with two replicate spots per field on NHS activated hydrogel
glass slides (CodeLink slides, Surmodics) using a contact-free piezoelectric microarray spotter
(SciFLEXARRAYER S3, Scienion, Berlin, Germany) equipped with one type 4 coated Piezo
Dispense Capillary (PDC80). Relative humidity was kept constant at 40% during the entire
print run. After printing, slides were incubated for 16 to 24 h in a humidity saturated chamber.
Remaining reactive NHS esters were subsequently quenched by incubation in 100 mM
ethanolamine, 50 mM sodium phosphate pH 9 for 1 h at room temperature (RT). Slides were
washed twice with water and dried by centrifugation at 300 x g using the CombiSlide system
(Eppendorf). Slides were stored dry at 4 °C until blocking.
Plant lectins
Slides were blocked for 30 min at RT with plant lectin binding buffer (1% (w/v) bovine serum
albumin (BSA, VWR Product ID 422361V) in 20 mM Hepes, 2 mM calcium chloride, 2 mM
magnesium chloride, 0.1 mM manganese chloride pH 7.0), rinsed with water and dried by
centrifugation. A 16 well incubation grid (FlexWell Incubation chamber, Grace BioLabs, Bend,
OR, USA) was attached to the surface. Fluorescein isothiocyanate (FITC) labeled plant lectins
(see SI Appendix Table S1; obtained from Vector Laboratories, Burlingame, CA, USA) were
diluted in plant lectin binding buffer. The diluted solutions (100 µL per well) were applied to the
slides and incubated overnight at 4 °C. Wells were washed for 3 min with plant lectin binding
buffer, the multiwell grid was removed, and the whole slide washed for 3 min with 10 mM
Hepes, 1 mM calcium chloride pH 7.0. After rinsing with water and drying by centrifugation,
slides were scanned using a GenePix 4300A microarray scanner (Molecular Devices,
Sunnyvale, CA, USA) at a detector voltage that prohibits detector signal saturation and at
10 µm resolution that is adequate for the size of the spots. Binding analysis: binding was
considered positive when a signal-to-noise ratio (SNR) of 7 was reached for at least three of
four spots from two independent experiments when the circle diameter was manually adjusted
to the spot size. Intensity evaluation: intensities were evaluated using GenePix Pro 7.2
(Molecular Devices) with a spot diameter of 70 μm at the position of greatest intensity, even if
S3
spots appeared larger, to minimize effects of spot size and subtraction of the intensity of the
spots printed with the printing buffer as background correction; these buffer subtracted values
are the “absolute” values given in the corresponding worksheet of the Supporting Data 3 Excel
workbook file. For each experimental spot, the absolute value was normalized to the highest
signal for the respective lectin. For example, the very first absolute binding intensity value for
lectin BSL for glycan “GA000005” listed in the worksheet is 77 (slide 1, spot 1), and this value
was normalized to the average of the best BSL binder “GA000175” spot intensities on the
same slide (77/(0.5*(47434+44265))=0.001679). All such resulting “relative” intensities are
given in the corresponding worksheet of the Supporting Data 3 Excel workbook file. The
reported value is generally the mean of four spots from two independent experiments unless
visibly unfit spots had to be removed from the analysis.
Monoclonal antibodies
Monoclonal rat antibody LM10 against oligoxylans (cell culture supernatant diluted 1:20,
PlantProbes, Leeds, UK)(1), murine monoclonal antibodies (concentration: 1 µg/mL, purity
>95% estimated by SDS-PAGE) 1A5(2), 1A10(3), and H16 (4) diluted in 3% BSA-PBS were
applied to blocked (1% BSA in 50 mM Hepes and 5 mM calcium chloride, pH 7.0, 1h at room
temperature) slides in 16 well format. Slides were incubated for 1 h at RT and washed three
times with PBS containing 0.1% Tween-20 (PBS-T). Secondary antibodies were diluted in 3%
BSA-PBS and incubated on the slide for 30 min. The following secondary antibodies were
used: goat anti-mouse IgG H+L AlexaFluor 635 (Life Technologies, Product-ID: A31574)
1:400; goat anti-rat IgG H+L AlexaFluor 555 (Life Technologies; Product-ID: A21434) 1:200.
Slides were washed once with PBS-T, twice with PBS, rinsed with water after removal of the
multiwell grid and dried by centrifugation. Intensities were read out using a GenePix 4300A
microarray scanner (Molecular Devices). Positive binding was evaluated from visual inspection
of the scanned glycan array images (2–5).
Sera from healthy human individuals
Sera for human studies were obtained as a gift from Sphingotec GmbH, Henningsdorf,
Germany. Sera from 15 apparently healthy human individuals were diluted 1:100 in 3% BSA-
PBS containing 0.1% Tween-20 and incubated at 37 °C for 20 min to dissolve lipid particles(6).
S4
Following centrifugation at 11000 rpm for 1 min, 100 µL of supernatant per serum sample was
applied to blocked slides (1% BSA-PBS, 30 min room temperature) with attached 16 well
incubation grid and incubated for 1.5 h at RT. Wells were washed three times for 5 min with
PBS-T. Secondary antibody solution (goat anti-human IgG-Fc AlexaFluor 488 (Dianova,
Product-ID: 109-545-098) diluted 1:400 in 3% BSA-PBS containing 0.1% Tween-20 was
applied to the wells. After incubation for 30 min at RT, slides were washed twice for 5 min with
PBS-T after which the multiwell gasket was removed and the slides washed in a Petri dish for
5 min with PBS, rinsed with water and dried by centrifugation. Slides were scanned using a
GenePix 4300A microarray scanner (Molecular Devices) at a detector voltage that prohibits
detector signal saturation at 10 µm resolution that is adequate for the size of the spots. Binding
analysis: binding was considered positive when a signal-to-noise ratio (SNR) of 7 was reached
for at least three of four spots from two independent experiments when the circle diameter was
manually adjusted to the spot size. Intensity evaluation: intensities were evaluated using
GenePix Pro 7.2 (Molecular Devices) with a spot diameter of 70 μm at the position of greatest
intensity, even if spots appeared larger, to minimize effects of spot size and subtraction of the
intensity of the spots printed with the printing buffer as background correction; these buffer
subtracted values are the “absolute” values given in the corresponding worksheet of the
Supporting Data 2 Excel workbook file. For both experimental repeats, the values were
normalized to the average intensity of all spots on the array. For example, the absolute value
for serum 1, slide 1, spot 1 is 2.5, which was divided by the average signal over all spots from
the first experiment (1658.39) to give the relative intensity of 1,51E-03. All such resulting
“relative” intensities are given in the corresponding worksheet of the Supporting Data 2 Excel
workbook file. The reported value is the mean of four spots from two glycan array experiments.
DC-SIGN
Generation and Expression of DC-SIGN-D
DC-SIGN-D was generated as a fusion protein consisting of the ECD of DC-SIGN and the Fc
part of human IgG1. Cloning and production was performed as described previously (7). In
brief, the sequence encoding the CRD and the neck domain of DC-SIGN (amino acids 61-404)
was amplified by RT-PCR and ligated in-frame into the expression vector pFUSE hIgG1-Fc2
(Invivogen, San Diego, CA, USA). The resulting construct was expressed in CHO-S cells using
the Freestyle™ MAX CHO Expression System (Life Technologies, Carlsbad, CA, USA).
Secreted protein was purified from the supernatant using a HiTrap™ Protein G HP column
S5
(GE Healthcare, Freiburg, Germany). Identity and homogeneity were confirmed by SDS-PAGE
and Western Blot analysis.
Expression of DC-SIGN-T
DC-SIGN-T (tetrameric ECD) comprising residues 66–404 was overexpressed in E. coli as
inclusion bodies, then refolded and purified to homogeneity as determined by SDS-PAGE and
size-exclusion chromatography in a functional form by an automated multistep purification
protocol using a high-throughput Akta Xpress System. This automated purification process,
relying on a mannan-agarose column, followed the report of Tabarani et al 2009(8).
Glycan arrays
Slides were blocked with 1% BSA (w/v) in 50 mM Hepes and 5 mM calcium chloride, pH 7.0
(BDC-buffer) for 1 h at rt, washed twice with water and dried by centrifugation (300 x g, 5 min).
A 16 well incubation grid was attached to the microarray slide. Recombinant DC-SIGN
constructs were diluted to 25 ng/µL, 5 ng/µL, and 1 ng/µL in BDC buffer. EDTA (final
concentration 10 mM) was added to a 25 ng/µL sample of each DC-SIGN construct.
Recombinant human IgG1-Fc was diluted to 25 ng/µL. All samples were incubated for 15 min
at 37°C to allow for calcium to dissociate in the negative controls. The samples were applied
to the glycan array slides (60 µL per well) and incubated at 4 °C for 18 h at high humidity and
washed three times with 0.1% Tween-20 (v/v) in 50 mM Hepes and 5 mM CaCl2, pH 7.0 (WDC-
buffer). To detect DC-SIGN-T, microarray wells were incubated with primary murine anti-
human CD209 (DC-SIGN) (Clone 9E9A8, Biolegend, Product-ID 330102) antibody diluted 1:50
in BDC-buffer for 30 min at RT, washed three times with WDC-buffer, and incubated for 30 min
with goat anti-mouse IgG Alexa Fluor 635 (Life Technologies, Product-ID: A31574) diluted
1:400. To detect DC-SIGN-D and human IgG1-Fc, wells were incubated with goat anti-human
IgG Alexa Fluor 647 (Life Technologies, Product-ID A21445) diluted 1:400 in BDC-buffer for 1
h at RT. Wells were washed once with WDC-buffer, twice with 10 mM Hepes, 1 mM calcium
chloride, rinsed with deionized water and dried by centrifugation (300 x g, 5 min). Read-out of
fluorescence intensity signals was performed with a GenePix 4300A microarray scanner
(Molecular Devices) at a detector voltage that prohibits detector signal saturation at 10 µm
resolution that is adequate for the size of the spots and image analysis was performed with the
GenePix Pro 7 software (Molecular Devices). Intensity evaluation: intensities were evaluated
with a spot diameter of 70 μm at the position of greatest intensity, even if spots appeared
larger, to minimize effects of spot size and subtraction of the intensity of the spots printed with
S6
the printing buffer as background correction; these buffer subtracted values are the “absolute”
values given in the two corresponding worksheets (DC-SIGN-T and DC-SIGN-D) of the
Supporting Data 4 Excel workbook file. These values were normalized to best binder Ley
antigen 157 at 25 ng/µL. One example of how the “relative” intensity values were obtained
(given in the two corresponding worksheets (DC-SIGN-T and DC-SIGN-D) of the Supporting
Data 4 Excel workbook file) is the first value for DC-SIGN-T (-132.5), which was then divided
by the average of the two spot intensities for best binder Ley antigen 157 at 25 ng/µL
(“GA000157”) as follows: -132.5/(0.5*(38408.5+40271.5))=-0.00337. All such resulting
“relative” intensities are given in the corresponding worksheet of the Supporting Data 2 Excel
workbook file. All such resulting “relative” intensities are given in the two corresponding
worksheets (DC-SIGN-T and DC-SIGN-D) of the Supporting Data 4 Excel workbook file.
Reported intensities are the mean of four spots from two independent glycan array experiments
normalized to the intensity of Ley antigen 157 at 25 ng/µL.
Surface Plasmon Resonance of Immobilized DC-SIGN and Immobilized Di-Hep
Binding avidity of DC-SIGN-T towards disaccharide 178 was measured at 25 °C using a
Biacore T100 instrument (GE Healthcare). DC-SIGN-T samples were cooled down to 10 °C
before injection into flow cells. For immobilization of oligosaccharides a C1 sensor chip surface
was modified with O-(2-aminoethyl)-O′-(2-carboxyethyl)-polyethylene glycol hydrochloride
(HCl·H2N-PEG-COOH) (Sigma Aldrich, molecular weight = 5,000). Sample and reference flow
cells were functionalized using 1 mg/mL HCl·H2N-PEG-COOH dissolved in 50 mM sodium
phosphate buffer, pH 8.5 at a flow rate of 10 μL/min and 420 s contact time using the Amine
Coupling Kit (GE Healthcare) according to the manufacturer’s recommendations, yielding
about 50 response units (RU). In the sample cell disaccharide 178 was immobilized at 1 mM
in 50 mM sodium phosphate buffer, pH 8.5 at a flow rate of 10 μL/min and 420 s contact time
using the Amine Coupling Kit (GE Healthcare), yielding about 140 RU. As a reference 1 mM
mono-Rha 167 was used under the same conditions, yielding about 9 RU. DC-buffer was used
as running buffer.
Binding runs were performed with DC-SIGN construct dilutions at indicated concentrations at
a flow rate of 30 μL/min for 60 s of contact time and 180 s of dissociation time. Flow cells were
regenerated twice with 2 M guanidine-hydrochloride for 30 s, followed by 2 M guanidine-
hydrochloride for 20 s and 300 s stabilization period. The binding signals were calculated as
the difference of RUs between ligand and reference flow cells and monitored as a function of
time represented as sensorgrams. Kinetic evaluation of binding responses was performed with
S7
the Biacore T100 Evaluation software (GE Healthcare), using reference-subtracted
sensorgrams. Apparent Kd values were determined with a steady-state affinity model.
Burkholderia lectins
Proteins BC2L-A and BC2L-C-ct were produced in E. coli BL21(DE3) as previously described
(9, 10). Transformed cells were cultured in LB medium (Duchefa Biochemie) containing
ampicillin (100 µg/mL) at 37 °C. When the culture reached an OD600 of 0.5, protein
overexpression was induced by isopropyl 1-thio-β-D-galactopyranoside (IPTG, Duchefa
Biochemie) added to a final concentration of 0.5 mM. Cells were incubated at 27 °C for 4 hours
(BC2L-A) or at 30 °C for 3 h (BC2L-C-ct), harvested by centrifugation and resuspended in
equilibration buffer (20 mM Tris/HCl, 100 mM NaCl, pH 7.4). Cells were then lysed by
sonication and cytosolic fractions containing soluble proteins were separated by centrifugation.
Lectins were purified by affinity chromatography on a D-mannose-agarose (Sigma-Aldrich)
column using an FPLC system (ÄKTA, GE Healthcare) and eluted with 20 mM Tris/HCl, 150
mM NaCl, 10 mM EDTA, pH 7.4. No impurities were detected using SDS-PAGE. Purified
proteins were dialysed against 10 mM Tris/HCl, 50 mM NaCl, 1 mM CaCl2 for 2 days, then
against 50 mM NH4HCO3 for 2 days, freeze-dried and stored at -20 °C
Freeze-dried lectins were dissolved in 10 mM HEPES, 150 mM NaCl, 0.005% TWEEN 20, pH
7.4 and incubated overnight at 4°C. Lectins were filtered and then transferred into conjugation
buffer (50 mM borate buffer, pH 8.5) using Zeba Spin Desalting Columns (Thermo Fisher
Scientific). Lectins in conjugation buffer were mixed in a 1:20 molar ratio with FITC (Thermo
Fisher Scientific) which was freshly dissolved in N,N-dimethylformamide to a concentration of
5 mg/mL. The solution was incubated with mixing at 400 rpm for 2 hours in the dark at RT.
Excess FITC was removed by subsequent buffer exchange (Zeba Spin Desalting Columns)
and lectins were transferred to buffer 100 mM Tris/HCl, 0.5 mM CaCl2, pH 7.4 during this
procedure. Lectins were concentrated by Vivaspin500 centrifugal concentrators (Sartorius
Stedim Biotech SA) to concentrations 0.8 mg/mL of BC2L-A and 1 mg/mL of BC2L-C-ct.
Activity of labelled lectins was tested by yeast agglutination experiment. 10 μL of BC2L-A (7.7
mg/ml) or BC2L-C-ct (10.5 mg/mL) was mixed with 10 μL of 5% yeast cell suspension and
incubated for 10 minutes at room temperature. Then the mixture was applied to a glass slide
and observed under fluorescent microscope (Olympus Corporation). Both lectins were active
because a well-visible agglutination was observed in the samples. Also, fluorescence of
agglutinates was visible in both samples. 10 μL of the buffer (100 mM Tris.Cl, 0.5 mM CaCl2,
S8
pH 7.42) instead of the lectin sample was used as a negative control and no agglutination was
observed in the sample.
Isothermal titration calorimetry using Auto-iTC200 (Malvern Instruments) was performed with
labelled lectins (dissolved in 100 mM Tris/HCl, 0.5 mM CaCl2, pH 7.4) to test the influence of
the label to their binding properties. 20 aliquots of 2 μL of methyl α-D-mannopyranoside
(dissolved in the same buffer) were automatically added at 4 minute interval to the protein
solution present in the calorimeter cell tempered to 25 ºC. Integrated heat effects were
analyzed using Microcal Origin 7 software (Malvern Instruments). Measurements were
performed in triplicates. Labelling had no significant influence on activity of the lectins because
obtained binding parameters corresponded to thermodynamic values of non-labelled lectins
(9, 10).
For glycan array incubation, lectins were diluted in 100 mM Tris/HCl pH 7.4 containing 2 mM
calcium chloride and 0.05% (v/v) Tween-20. The concentration series of the lectins differed
slightly resulting in the 1 mg/mL sample shown in Figure 6 to be 0.96 mg/mL for BC2L-A and
1.05 mg/mL for BC2L-C-ct. At this concentration, EDTA was added to a final concentration of
5 mM as negative binding control. Lectins were incubated for 15 min at 37 °C to allow for
calcium to dissociate in the negative controls and then applied to unblocked slides (60 µL per
well). Microarrays were incubated for 1 h at RT. Wells were washed twice for 1 min with 20
mM Tris/HCl pH 7.4. The incubation grid was removed; the slides washed for 30 s with water,
dried by centrifugation and scanned with a GenePix 4300A microarray scanner (Molecular
Devices) at a detector voltage that prohibits detector signal saturation at 10 µm resolution that
is adequate for the size of the spots. Intensity evaluation: intensities were quantified using
GenePix Pro 7.2 (Molecular Devices). To adjust for different spots sizes, spot intensities were
evaluated from 70 µm diameter circles in the center of the spots and the values for the printing
buffer spots from the same well were subtracted as background correction; these buffer
subtracted values are the “absolute” values given in the two corresponding worksheets (BC2L-
A and BC2L-C-ct) of the Supporting Data 1 Excel workbook file. For every slide, intensities
were normalized to the signal of the same lectin for arabinomannan hexasaccharide 257 at
962.5 µg/µL or 1050 µg/µL, closest to a 1 mg/mL concentration. One example calculation
follows for the normalization of the first absolute value given in the table for BC2L-A at 1925
µg/µL, slide 1, spot 1 (53.5): 53.5/(0.5+(46214+46150))=0.01158. All such resulting “relative”
intensities are given in the two corresponding worksheets (BC2L-A and BC2L-C-ct) of the
Supporting Data 1 Excel workbook file. The reported values are generally the mean of six spots
from three independent experiments unless intensities had to be removed because of aberrant
spot shapes. For avidity analysis, fluorescence intensities (generally the mean of two spots
per glycan, lectin and concentration for three concentration series) of glycans containing an
S9
aminopentanol linker were plotted against lectin concentration using SigmaPlot 13.
Background-corrected intensities (I) were fitted against the hyperbolic equation 𝐼 =
𝐼𝑚𝑎𝑥𝑐(𝑃𝑟𝑜𝑡𝑒𝑖𝑛)
𝐾𝑑+𝑐(𝑃𝑟𝑜𝑡𝑒𝑖𝑛) with Kd being the apparent dissociation constant.
Diversity scores
To quantify the diversity of the glycan libraries we used the glycan fingerprint method as
previously described (11). Briefly, glycans were converted into xml format and 512 bit
fingerprints were calculated form all possible sub-trees of a glycan structure up to a chain
length of seven. The resulting hashed bit strings were then compared using the modified
Tanimoto coefficient to yield a similarity score between zero and one. The diversity of each
library was calculated from the average pairwise distance of all members. This matrix was also
used to build the tree using a neighbor-joining algorithm(12) and FigTree (version 1.4.3,
http://tree.bio.ed.ac.uk/software/figtree/) for visualization.
S10
2. Supporting Tables
Table S1: Plant lectins used for validation of glycan printing. Colors in specificity column
correspond to the colors used in Figure 3 and SI Appendix Figure S1. Specificity assignment
is based on vendor data and review publications(13, 14, 15). Additional literature precedent is
indicated for specificities not stated in these general sources.
Lectin Conc. Specificity
Bandeiraea simplicifolia lectin I (BSL) 20 µg/mL Orange: Terminal α-Gal
Blue: Terminal α-GalNAc
Concanavalin A (ConA) 20 µg/mL Orange: Terminal α-Man
Blue: Terminal α-Glc
Dolichos biflorus agglutinin (DBA) 50 µg/mL Orange: Terminal α-GalNAc
Blue: Terminal β-GalNAc
Erythrina cristagalli lectin (ECL) 20 µg/mL Orange: Terminal β-Gal
Maackia amurensis lectin I (MALI) 20 µg/mL Orange: Terminal β-Gal(1,4)Glc/GlcNAc
Blue: Terminal α-Neu5Ac(2,3)β-Gal(1,4)Glc/GlcNAc
(Both cases except glycans with fucose linked to
GlcNAc) (16)
Peanut agglutinin (PNA) 20 µg/mL Orange: Terminal β-Gal
Ricinus communis agglutinin
(RCA120)
20 µg/mL Orange: Terminal β-Gal
Blue: Terminal α-Neu5Ac(2,6)β-Gal (18)
Soybean agglutinin (SBA) 20 µg/mL Orange: Terminal GalNAc
Blue: Terminal Gal
Sambucus nigra lectin (SNL) 50 µg/mL Orange: Terminal α-(2,6) sialic acid
Ulex europaeus agglutinin I (UEAI) 50 µg/mL Orange: Terminal α-Fuc
Wheat germ agglutinin (WGA) 20 µg/mL Orange: Terminal GlcNAc or sialic acid
Blue: Terminal GalNAc (19, 20)
S11
Table S2: Glycans immobilized on 140 compound array.
Glycan ID
Glycan Structure (Microbial origin if applicable)
Reference DOI
Lectin Detected by mono-
clonal antibody
Signal from n human
sera (out of 15)
Number saccharide
units
Printed in
Predicted Bound (of predicted)
Not Predicted
Bounc
Predicted Not Bound
5
Neu5Ac(a2-6)Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminohexanol
Hanashima (2007)
10.1002/asia.200600424 RCA, SNL,
WGA RCA, SNL WGA >5 5 NaP 8.5
6
Neu5Ac(a2-3)Gal(b1-3)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminohexanol
Hanashima (2007)
10.1002/asia.200600424 WGA WGA >5 5 NaP 8.5
7
Fuc(a1-3)[Neu5Ac(a2-3)Gal(b1-4)]GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminohexanol
Hanashima (2007b)
10.1021/ol0704946 UEA, WGA WGA UEA 3 6 NaP 8.5
8
Neu5Ac(a2-6)Gal(b1-4)Glc(b1-1)aminohexanol
Hanashima (2007)
10.1002/asia.200600424 RCA, SNL,
WGA RCA, SNL,
WGA 3 NaP 8.5
9
Neu5Ac(a2-3)Gal(b1-4)Glc(b1-1)aminohexanol
Hanashima (2007)
10.1002/asia.200600424 MAL I, WGA MAL I, WGA 2 3 NaP 8.5
S12
10
Neu5Ac(a2-6)Gal(b1-4)GlcNAc-6-sulfate(b1-1)aminohexanol
RCA, SNL,
WGA RCA, SNL,
WGA ECL 1 3 NaP 8.5
11
Gal(b1-4)Glc(b1-1)aminohexanol
ECL, MAL I, PNA, RCA,
SBA
ECL, MAL I, PNA, RCA,
SBA >5 2 NaP 8.5
12
Gal(b1-4)GlcNAc-6-sulfate(b1-1)aminohexanol
ECL, MAL I, PNA, RCA,
SBA
ECL, MAL I, RCA, SBA
WGA PNA >5 2 NaP 8.5
69
Araf(a1-5)Araf(a1-1)aminopentanol
(Mycobacterium tuberculosis)
Kandasamy (2013)
10.1039/c3cc00042g >5 2 NaP 8.5
70
Araf(a1-5)Araf(a1-3)[Araf(a1-5)Araf(a1-5)]Araf(a1-5)Araf(a1-1)aminopentanol
(Mycobacterium tuberculosis)
Kandasamy (2013)
10.1039/c3cc00042g >5 6 NaP 8.5
S13
71
Araf(a1-3)[Araf(a1-5)]Araf(a1-1)aminopentanol
(Mycobacterium tuberculosis)
Kandasamy (2013)
10.1039/c3cc00042g >5 3 NaP 8.5
72
Araf(a1-5)Araf(a1-5)Araf(a1-5)Araf(a1-5)Araf(a1-5)Araf(a1-5)aminopentanol
(Mycobacterium tuberculosis)
Kandasamy (2013)
10.1039/c3cc00042g >5 6 NaP 8.5
73
Col(a1-3)[Col(a1-6)]Glc(a1-4)Gal(a1-3)GlcNAc(b1-1)aminopentanol
(Escherichia coli O111)
Calin (2013) 10.1002/chem.20120439
4 >5 5 NaP 8.5
74
ManNAc(b1-3)FucNAc(a1-3)GalNAc(a1-4)Gal(a1-1)aminopentanol
(Streptococcus pneumoniae)
Pereira (2015) 10.1002/anie.201504847 BSL H16 >5 4 NaP 8.5
75
GalNAc(a1-4)Gal(a1-1)aminopentanol
(Streptococcus pneumoniae)
Pereira (2015) 10.1002/anie.201504847 BSL, DBA,
SBA, WGA BSL, SBA,
WGA SNL DBA H16 >5 2 NaP 8.5
S14
76
GalNAc(b1-4)Gal(a1-1)aminopentanol
Pereira (2015) 10.1002/anie.201504847 DBA, SBA,
WGA DBA, SBA,
WGA BSL >5 2 NaP 8.5
77
FucNAc(a1-3)GalNAc(a1-4)Gal(a1-1)aminopentanol
(Streptococcus pneumoniae)
Pereira (2015) 10.1002/anie.201504847 BSL, UEA >5 3 NaP 8.5
78
FucNAc(b1-3)GalNAc(a1-4)Gal(a1-1)aminopentanol
Pereira (2015) 10.1002/anie.201504847 BSL, SBA, UEA
H16 >5 3 NaP 8.5
80
GalNAc(b1-1)aminoethanol
Pereira (2015) 10.1002/anie.201504847 DBA, SBA,
WGA DBA, SBA,
WGA BSL, RCA,
SNL >5 1 NaP 8.5
81
FucNAc(a1-1)aminopentanol
Lisboa (2017) 10.1073/pnas.170687511
4 UEA H16 >5 1 NaP 8.5
82
Man(a1-2)Man(a1-2)[Gal(b1-4)]Man(a1-1)aminopentanol
(Leishmania donovani)
Anish (2013) 10.1021/cb400602k ConA, ECL, PNA, RCA,
SBA
ConA, ECL, PNA, RCA,
SBA >5 4 NaP 8.5
S15
83
Man(a1-2)Man(a1-2)Man(a1-1)aminopentanol
(Leishmania donovani)
ConA ConA >5 3 NaP 8.5
84
Gal(b1-4)Man(a1-1)aminopentanol
(Leishmania chagasi)
Anish (2013) 10.1021/cb400602k ECL, PNA, RCA, SBA
ECL, PNA, RCA, SBA
ConA >5 2 NaP 8.5
85
Man(a1-2)Man(a1-1)aminopentanol
ConA ConA 2 2 NaP 8.5
90
Glc(b1-1)aminoethanol
Parameswarappa (2016)
10.1016/j.chembiol.2016.09.016
>5 1 NaP 8.5
91
GlcNAc(a1-2)Hep(a1-3)Hep(a1-5)Kdo(a2-1)aminopentanol
(Neisseria meningitidis)
Yang (2012) 10.1039/c1sc00804h;
10.1021/ja401164s WGA WGA 1A5 >5 4 NaP 8.5
92
Hep(a1-3)Hep(a1-5)Kdo(a2-1)aminopentanol
(Neisseria meningitidis)
Yang (2013) 10.1021/ja401164s 1A5 3 NaP 8.5
S16
93
Hep(a1-3)Hep(a1-5)[L-Ara4N(b1-8)]Kdo(a2-1)aminopentanol
(Proteus spec.)
Yang (2013) 10.1021/ja401164s 1A5 1 4 NaP 8.5
94
Hep(a1-7)Hep(a1-3)Hep(a1-5)Kdo(a2-1)aminopentanol
(Yersinia pestis)
Yang (2013) 10.1021/ja401164s 1A5 4 NaP 8.5
95
Hep(a1-2)Hep(a1-3)Hep(a1-5)Kdo(a2-1)aminopentanol
(Haemophilus influenzae)
Yang (2013) 10.1021/ja401164s 1A5 4 NaP 8.5
96
Hep(a1-5)Kdo(a2-1)aminopentanol
Yang (2013) 10.1021/ja401164s BSL 1A5 2 NaP 8.5
97
Hep(a1-7)Hep(a1-3)Hep(a1-1)aminopentanol
(Yersinia pestis)
Anish (2013b) 10.1002/anie.201301633 3 NaP 8.5
S17
98
Kdo(a2-8)Kdo(a2-4)Kdo(a2-1)aminopentanol
(Chlamydia spec.)
1A5 >5 3 NaP 8.5
99
Kdo(a2-1)aminopentanol
Yang (2013) 10.1021/ja401164s 1A5 >5 1 NaP 8.5
100
Hep(a1-1)aminopentanol
Yang (2013) 10.1021/ja401164s 3 1 NaP 8.5
101
Glc(b1-1)aminopentanol
Leonori (2013) 10.3762/bjoc.9.38 >5 1 NaP 8.5
102
D-FucNAc(b1-1)aminopentanol
Leonori (2013) 10.3762/bjoc.9.38 BSL, DBA, RCA, SNL,
WGA >5 1 NaP 8.5
103
FucNAc(b1-1)aminopentanol
Leonori (2013) 10.3762/bjoc.9.38 UEA H16 >5 1 NaP 8.5
104
Glc(b1-3)D-FucNAc(b1-1)aminopentanol
(Pseudomonas aeruginosa)
Leonori (2013) 10.3762/bjoc.9.38 >5 2 NaP 8.5
S18
105
Glc(b1-3)FucNAc(b1-1)aminopentanol
(Pseudomonas aeruginosa)
Leonori (2013) 10.3762/bjoc.9.38 >5 2 NaP 8.5
153
Gal(b1-3)GalNAc(a1-1)aminopentanol
ECL, PNA, RCA, SBA
PNA, RCA, SBA
ECL >5 2 NaP 8.5
154
Fuc(a1-3)[Gal(b1-4)]GlcNAc(b1-1)aminopentanol
Schlegel (2011) 10.1002/cbic.201100511 ECL, PNA, RCA, SBA,
UEA
BSL ECL, PNA, RCA, SBA,
UEA 2 NaP 8.5
155
Neu5Ac(a2-6)GalNAc(a1-1)aminopentanol
SNL, WGA WGA BSL, PNA, RCA, SBA
SNL 4 2 NaP 8.5
156
Gal(b1-4)[Gal(b1-4)Glc(b1-6)]GlcNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
ECL, MAL I, PNA, RCA,
SBA
ECL, MAL I, PNA, RCA,
SBA WGA >5 4 NaP 8.5
S19
157
Fuc(a1-3)[Fuc(a1-2)Gal(b1-4)]GlcNAc(b1-1)aminopentanol
UEA UEA BSL 4 NaP 8.5
158
Gal(b1-3)[Fuc(a1-4)]GlcNAc(b1-1)aminopentanol
ECL, PNA, RCA, SBA,
UEA UEA
ECL, PNA, RCA, SBA
1 3 NaP 8.5
159
Fuc(a1-2)Gal(b1-3)[Fuc(a1-4)]GlcNAc(b1-1)aminopentanol
UEA UEA BSL 2 3 NaP 8.5
160
Gal-2,3-Pyruvate(a1-1)aminopentanol (mixture of R/S pyruvate)
(Streptococcus pneumoniae)
Pereira (2015) 10.1002/anie.201504847 BSL >5 1 NaP 8.5
161
Gal(a1-3)Gal(b1-4)Glc(b1-1)aminopentanol
Hofmann (2015) 10.1038/nature15388;
10.1021/acs.joc.6b00554 BSL, SBA BSL, SBA RCA >5 3 NaP 8.5
162
Gal(a1-3)Gal(b1-4)GlcNAc(b1-1)aminopentanol
Hahm (2016) 10.1021/acs.joc.6b00554 BSL, SBA BSL, SBA ECL, RCA,
WGA >5 3 NaP 8.5
S20
163
Gal(a1-3)Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
Hahm (2016) 10.1021/acs.joc.6b00554 BSL, SBA BSL, SBA RCA, WGA >5 5 NaP 8.5
164
Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
(Streptococcus pneumoniae)
Hahm (2016) 10.1021/acs.joc.6b00554 ECL, MAL I, PNA, RCA,
SBA
ECL, MAL I, RCA, SBA
WGA PNA >5 4 NaP 8.5
165
Fuc(a1-2)Gal(b1-3)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
Hahm (2016) 10.1021/acs.joc.6b00554 UEA UEA >5 5 NaP 8.5
166
Gal(b1-3)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
Hahm (2016) 10.1021/acs.joc.6b00554 ECL, PNA, RCA, SBA
PNA, RCA, SBA
ECL >5 4 NaP 8.5
167
Rha(a1-1)aminopentanol
Martin (2013) 10.1021/ja401410y 1A10 >5 1 NaP 8.5
168
Rha(a1-3)Glc(b1-1)aminopentanol
Martin (2013) 10.1021/ja401410y 1A10 >5 2 NaP 8.5
169
Glc(a1-2)Glc(a1-1)aminopentanol
(Clostridium difficile)
Martin (2013) 10.1021/ja401410y ConA ConA >5 2 NaP 8.5
S21
170
Glc(b1-4)Glc(a1-2)Glc(a1-1)aminopentanol
(Clostridium difficile)
Martin (2013) 10.1021/ja401410y ConA >5 3 NaP 8.5
171
Rha(a1-3)Glc(b1-4)Glc(a1-1)aminopentanol
(Clostridium difficile)
Martin (2013) 10.1021/ja401410y 1A10 >5 3 NaP 8.5
172
Gal(b1-3)GalNAc(b1-3)Gal(a1-4)Gal(b1-4)Glc(b1-1)aminopentanol
ECL, PNA, RCA, SBA
PNA, RCA, SBA
BSL ECL >5 5 NaP 8.5
173
Neu5Ac(a2-8)Neu5Ac(a2-3)[GalNAc(b1-4)]Gal(b1-4)Glc(b1-1)aminopentanol
DBA, SBA,
WGA DBA, SBA WGA 5 NaP 8.5
174
Gal(a1-4)Gal(b1-4)Glc(b1-1)aminopentanol
Hofmann (2015) 10.1038/nature15388 BSL, SBA BSL, SBA RCA >5 3 NaP 8.5
175
GalNAc(a1-1)AminoLinker2
Schlegel (2011) 10.1002/cbic.201100511 BSL, DBA, SBA, WGA
BSL, DBA, SBA, WGA
2 1 NaP 8.5
S22
176
Fuc(a1-3)[Gal(b1-4)]GlcNAc(b1-1)AminoLinker2
Schlegel (2011) 10.1002/cbic.201100511 ECL, PNA, RCA, SBA,
UEA
BSL ECL, PNA, RCA, SBA,
UEA 3 NaP 8.5
177
GlcNAc(a1-2)Hep(a1-3)Hep(a1-1)aminopentanol
(Neisseria meningitidis)
Reinhardt (2015) 10.1016/j.chembiol.2014.
11.016 WGA WGA >5 3 NaP 8.5
178
Hep(a1-3)Hep(a1-1)aminopentanol
Reinhardt (2015) 10.1016/j.chembiol.2014.
11.016 1 2 NaP 8.5
179
Gal(b1-4)Glc(b1-1)aminopentanol
ECL, MAL I, PNA, RCA,
SBA
ECL, MAL I, PNA, RCA,
SBA >5 2 NaP 8.5
180
GalNAc(b1-4)Gal(b1-4)Glc(b1-1)aminopentanol
DBA, SBA,
WGA DBA, SBA WGA >5 3 NaP 8.5
181
Neu5Ac(a2-3)Gal(b1-4)Glc(b1-1)aminopentanol
Fair (2015) 10.1039/c5cc01368b MAL I, WGA MAL I, WGA 2 3 NaP 8.5
S23
182
GalNAc-4-sulfate(b1-1)aminopentanol
Kandasamy (2014)
10.1039/c3cc48860h 1 1 NaP 8.5
183
IdoA-2,4-disulfate(a1-1)aminopentanol
Nonaka (2014) 10.1039/c3cc48860h;
ConA, DBA, ECL, PNA, RCA, SNL,
UEA
>5 1 NaP 8.5
184
IdoA(a1-3)GalNAc-4-sulfate(b1-1)aminopentanol
Kandasamy (2014)
10.1039/c3cc48860h 2 NaP 8.5
185
IdoA-2-sulfate(a1-3)GalNAc-4-sulfate(b1-1)aminopentanol
Kandasamy (2014)
10.1039/c3cc48860h ConA, SNL 2 NaP 8.5
186
IdoA(a1-3)GalNAc(b1-1)aminopentanol
Kandasamy (2014)
10.1039/c3cc48860h 1 2 NaP 8.5
187
GlcA(b1-4)Glc(b1-3)GlcA(b1-4)Glc(b1-1)aminoethanol
(Streptococcus pneumoniae)
Parameswarappa (2016)
10.1016/j.chembiol.2016.09.016
>5 4 NaP 8.5
S24
188
Glc(b1-3)GlcA(b1-4)Glc(b1-1)aminoethanol
(Streptococcus pneumoniae)
Parameswarappa (2016)
10.1016/j.chembiol.2016.09.016
>5 3 NaP 8.5
189
GalNAc(a1-1)Thr-Linker
BSL, DBA, SBA, WGA
BSL, DBA, SBA, WGA
SNL 5 1 NaP 8.5
190
Glc(b1-3)Glc(b1-3)[Glc(b1-6)]Glc(b1-3)Glc(b1-1)aminopentanol
(Candida spec.)
Weishaupt (2017)
10.1039/C7CC00520B >5 5 NaP 8.5
191
Glc(b1-3)Glc(b1-3)[Glc(b1-6)]Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-1)aminopentanol
(Candida spec.)
Weishaupt (2017)
10.1039/C7CC00520B >5 9 NaP 8.5
192
Glc(b1-3)Glc(b1-3)[Glc(b1-6)]Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-1)aminopentanol
(Candida spec.)
Weishaupt (2017)
10.1039/C7CC00520B >5 13 NaP 8.5
193
Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-3)Glc(b1-1)aminopentanol
(Candida spec.)
Weishaupt (2013)
10.1002/chem.201204518
>5 12 NaP 8.5
S25
194
L-PneNAc(a1-2)GlcA(b1-3)FucNAc(a1-3)D-FucNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4 H16 >5 4 NaP 8.5
195
Mixture of: D-6d-xylHexpNAc-4-ulo(b1-1)aminopentanol (Sugp(b1-1)aminopentanol) and D-FucNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4
BSL, DBA, RCA, SNL,
WGA 3 1 NaP 8.5
196
Mixture of: FucNAc(a1-3)D-6d-xylHexpNAc-4-ulo(b1-1)aminopentanol and FucNAc(a1-3)D-FucNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4 H16 >5 2 NaP 8.5
197
FucNAc(a1-3)D-FucNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4 H16 >5 2 NaP 8.5
198
GlcA(b1-4)FucNAc(a1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4 H16 >5 2 NaP 8.5
S26
199
Glc(b1-3)FucNAc(a1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4 H16 >5 2 NaP 8.5
200
L-PneNAc(a1-2)GlcA(b1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4 5 2 NaP 8.5
201
L-PneNAc(a1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4 H16 5 1 NaP 8.5
202
L-PneNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
Lisboa (2017) 10.1073/pnas.170687511
4 >5 1 NaP 8.5
203
Gal(b1-4)[Glc(b1-6)]GlcNAc(b1-3)Gal(b1-1)aminopentanol
(Streptococcus pneumoniae)
ECL, MAL I, RCA, SBA
ECL, MAL I, RCA, SBA
WGA >5 4 NaP 8.5
204
Glc(a1-4)Gal(a1-4)GlcA(b1-4)Glc(b1-1)aminoethanol
(Streptococcus pneumoniae)
Schumann (2017) 10.1126/scitranslmed.aaf
5347 ConA ConA >5 4 NaP 8.5
S27
205
Glc(a1-4)Gal(a1-1)aminoethanol
(Streptococcus pneumoniae)
Schumann (2017) 10.1126/scitranslmed.aaf
5347 ConA ConA >5 2 NaP 8.5
206
GlcA(b1-4)Glc(b1-4)Glc(a1-4)Gal(a1-1)aminoethanol
(Streptococcus pneumoniae)
Schumann (2017) 10.1126/scitranslmed.aaf
5347 >5 4 NaP 8.5
207
Glc(a1-4)Gal(a1-4)GlcA(b1-4)Glc(b1-1)aminopentanol
(Streptococcus pneumoniae)
Schumann (2017) 10.1126/scitranslmed.aaf
5347 ConA ConA >5 4 NaP 8.5
208
Gal(a1-4)GlcA(b1-4)Glc(b1-4)Glc(a1-1)aminopentanol
(Streptococcus pneumoniae)
Schumann (2017) 10.1126/scitranslmed.aaf
5347 BSL, SBA BSL SBA >5 4 NaP 8.5
209
GlcA(b1-4)Glc(b1-4)Glc(a1-4)Gal(a1-1)aminopentanol
(Streptococcus pneumoniae)
Schumann (2017) 10.1126/scitranslmed.aaf
5347 >5 4 NaP 8.5
210
Glc(b1-4)Glc(a1-4)Gal(a1-4)GlcA(b1-1)aminopentanol
(Streptococcus pneumoniae)
Schumann (2017) 10.1126/scitranslmed.aaf
5347 >5 4 NaP 8.5
S28
211
Xyl(b1-4)Xyl(b1-1)aminopentanol
Schmidt (2015) 10.1002/chem.20150006
5 LM10 3 2 NaP 8.5
212
Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-1)aminopentanol
Schmidt (2015) 10.1002/chem.20150006
5 LM10 2 4 NaP 8.5
213 Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-1)aminopentanol
Schmidt (2015) 10.1002/chem.20150006
5 LM10 2 6 NaP 8.5
214 Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-1)aminopentanol
Schmidt (2015) 10.1002/chem.20150006
5 LM10 2 8 NaP 8.5
215
Glc(b1-4)Glc(b1-4)Glc(b1-4)Glc(b1-1)aminopentanol
PNA >5 4 NaP 8.5
216
Glc(b1-3)GlcA(b1-4)Glc(b1-1)aminopentanol
(Streptococcus pneumoniae)
Weishaupt (2016)
10.3762/bjoc.12.139 >5 3 NaP 8.5
217
GlcA(b1-4)Glc(b1-1)aminoethanol
(Streptococcus pneumoniae)
Parameswarappa (2016)
10.1016/j.chembiol.2016.09.016
>5 2 NaP 8.5
S29
218
Glc(b1-3)GlcA(b1-1)aminoethanol
(Streptococcus pneumoniae)
Parameswarappa (2016)
10.1016/j.chembiol.2016.09.016
>5 2 NaP 8.5
219
ManNAc(b1-3)FucNAc(a1-3)GalNAc(a1-4)Gal-2,3-pyruvate(a1-1)aminopentanol
(Streptococcus pneumoniae)
Pereira (2015) 10.1002/anie.201504847 H16 >5 4 NaP 8.5
220
GlcA(b1-1)aminoethanol
Parameswarappa (2016)
10.1016/j.chembiol.2016.09.016
1 NaP 8.5
225
Glc(a1-4)GalNAc(b1-4)Man(a1-1)aminopentanol
(Toxoplasma gondii)
Goetze (2014) 10.1002/anie.201406706 ConA ConA >5 3 NaP 8.5
226
Glc(a1-4)GalNAc(b1-4)[Man(a1-2)Man(a1-6)]Man(a1-1)aminopentanol
(Toxoplasma gondii)
Goetze (2014) 10.1002/anie.201406706 ConA ConA >5 5 NaP 8.5
S30
227
Glc(a1-4)GalNAc(b1-4)[Man-6-PEtN(a1-2)Man(a1-6)]Man(a1-1)aminopentanol
(Toxoplasma gondii)
Goetze (2014) 10.1002/anie.201406706 ConA ConA >5 5 NaP 8.5
229
GalNAc(b1-4)Man(a1-1)aminopentanol
DBA, SBA,
WGA DBA, SBA,
WGA BSL, ECL, RCA, SNL
>5 2 NaP 8.5
230
GalNAc(b1-4)[Man-6-PEtN(a1-2)Man(a1-6)]Man(a1-1)aminopentanol
DBA, SBA,
WGA DBA, SBA,
WGA ConA, ECL 4 4 NaP 8.5
231
GalNAc(b1-4)[Man(a1-2)Man(a1-6)]Man(a1-1)aminopentanol
ConA, DBA, SBA, WGA
ConA, DBA, SBA, WGA
ECL, RCA 4 4 NaP 8.5
S31
232
GalNAc(b1-4)[Man-6-PEtN(a1-2)Man(a1-6)]Man-2-PEtN(a1-1)aminopentanol
DBA, SBA,
WGA DBA, SBA ConA WGA 2 4 NaP 8.5
233
GalNAc(b1-4)Man(a1-1)aminododecanol
DBA, SBA,
WGA DBA, SBA,
WGA ECL >5 2 NaP 8.5
234
GalNAc(b1-4)Man(a1-1)p-aminocyclohexanol
DBA, SBA,
WGA DBA, SBA,
WGA ConA, ECL >5 2 NaP 8.5
235
GlcA(b1-4)Glc(b1-3)GlcA(b1-1)aminoethanol
(Streptococcus pneumoniae)
Parameswarappa (2016)
10.1016/j.chembiol.2016.09.016
>5 3 NaP 8.5
236
GlcA(b1-4)Glc(b1-3)Glc(b1-4)Glc(b1-1)aminoethanol and/or Glc(b1-4)Glc(b1-3)GlcA(b1-4)Glc(b1-1)aminoethanol
>5 4 NaP 8.5
237
Man(a1-1)aminopentanol
ConA ConA 1 NaP 8.5
S32
238
GlcNAc-6-P-phosphoaminopentanol(a1-3)GlcNAc-6-P-phosphoaminopentanol(a1-2)glyceric acid
(Clostridium difficile)
Martin (2013b) 10.1039/c3cc43545h >5 2 NaP 8.5
239
GlcA(a1-3)Gal(a1-3)ManNAc(b1-4)Glc(b1-4)Glc(a1-1)aminopentanol
(Streptococcus pneumoniae)
>5 5 NaP 8.5
240
GlcA(a1-3)Gal(a1-3)ManNAc-6-acetate(b1-4)Glc(b1-4)Glc(a1-1)aminopentanol
(Streptococcus pneumoniae)
5 5 NaP 8.5, PBS
241
GlcA(a1-3)Gal(a1-1)aminopentanol
(Streptococcus pneumoniae)
>5 2 NaP 8.5
242
Glc(b1-4)Glc(a1-1)aminopentanol
PNA >5 2 NaP 8.5
243
ManNAc(b1-4)Glc(b1-4)Glc(a1-1)aminopentanol
(Streptococcus pneumoniae)
>5 3 NaP 8.5
S33
244
GalNAc(b1-3)GalNAc(b1-1)aminopentanol
DBA, SBA,
WGA SBA SNL DBA, WGA 3 2 NaP 8.5
245
Glc(b1-4)Gal(b1-4)Glc(b1-1)aminopentanol
Hofmann (2015) 10.1038/nature15388 RCA, SBA >5 3 NaP 8.5
247
Rha(a1-3)[Rha(a1-3)Glc(b1-4)]Glc(a1-2)Glc(a1-1)aminopentanol
(Clostridium difficile)
Martin (2013) 10.1021/ja401410y ConA 1A10 >5 5 NaP 8.5
248
GlcNAc(a1-3)GlcNAc-6-P-phosphoaminopentanol(a1-2)glyceric acid
(Clostridium difficile)
Martin (2013b) 10.1039/c3cc43545h WGA WGA >5 2 NaP 8.5
249
GlcNAc(a1-3)GlcNAc[(a1-2)glyceric acid](6-P-6)GlcNAc(a1-3)GlcNAc-6-P-phosphoaminopentanol(a1-2)glyceric acid
(Clostridium difficile)
Martin (2013b) 10.1039/c3cc43545h WGA WGA >5 4 NaP 8.5
S34
250
Man(a1-2)Man(a1-2)[Gal(b1-4)]Man(a1-1)aminoethanol
(Leishmania donovani)
Anish (2013) 10.1021/cb400602k ConA, ECL, PNA, RCA,
SBA
ConA, ECL, PNA, RCA
SBA >5 4 NaP 8.5
251
Glc(b1-3)Gal(b1-4)Man(a1-1)aminopentanol
(Leishmania chagasi)
Anish (2013) 10.1021/cb400602k 3 3 NaP 8.5
252
Rha(a1-2)Rha(a1-2)Rha(a1-1)aminopentanol
(Klebsiella pneumoniae)
Seeberger (2017) 10.1002/anie.201700964 1A10 >5 3 NaP 8.5
253
GalNAc-2,3-Oxazolidinone(a1-4)GalNAc-2,3-Oxazolidinone(a1-1)aminopentanol
BSL, SBA, SNL, WGA
>5 2 NaP 8.5
S35
254
Glc(a1-2)Glc(a1-3)[FucNAc(a1-3)GalNAc(b1-4)]ManNAcA(b1-1)aminopentanol
(Streptococcus pneumoniae)
Seeberger (2017b)
10.3762/bjoc.13.19 ConA ConA H16 >5 5 NaP 8.5
255
Glc(a1-2)Glc(a1-3)[Gal(a1-3)FucNAc(a1-3)GalNAc(b1-4)]ManNAcA(b1-1)aminopentanol
(Streptococcus pneumoniae)
Seeberger (2017b)
10.3762/bjoc.13.19 BSL, ConA,
SBA BSL ConA, SBA H16 >5 6 NaP 8.5
256
Araf(a1-3)[Araf(a1-5)]Araf(a1-5)Araf(a1-1)aminopentanol
(Mycobacterium tuberculosis)
>5 4 NaP 8.5
257
Man(a1-5)Araf(a1-3)[Man(a1-5)Araf(a1-5)]Araf(a1-5)Araf(a1-1)aminopentanol
(Mycobacterium tuberculosis)
Hahm (2017) 10.1073/pnas.170014111
4 ConA ConA >5 6 NaP 8.5
S36
258
GalNAc-3,4-diacetate(a1-4)GalNAc-3-acetate(a1-4)GalNAc-3-acetate(a1-4)GalNAc-3-acetate(a1-1)aminopentanol
DBA >5 4 PBS
Abbreviations: NaP 8.5 (50 mM sodium phosphate buffer, pH 8.5), PBS (phosphate buffered saline); For lectins see Supporting Table 1
S37
Monosaccharide representations:
S38
All synthetic glycans meet the standards of purity set by the Jounral of Organic Chemistry which means that 1H-, 13C-NMR as well as high resolution mass spec analyses were performed. The following publications describe the synthesis of the majority of glycans as referred to in the table.
References Anish (2013) ACS Chem. Biol. 8 2412−2422 (2013) 10.1021/cb400602k
Anish (2013b) Angew. Chem. Int. Ed. 52 9524 –9528 (2013) 10.1002/anie.201301633 Calin (2013) Chem. Eur. J. 19 3995 – 4002 (2013) 10.1002/chem.201204394 Fair (2015) Chem. Commun. 51 6183-6185 (2015) 10.1039/c5cc01368b Goetze (2014) Angew. Chem. Int. Ed. 53 13701 –13705 (2014) 10.1002/anie.201406706 Hahm (2016) J. Org. Chem. 81 5866–5877 (2016) 10.1021/acs.joc.6b00554 Hahm (2017) Proc .Natl. Acad. Sci. USA 114 E3385–E3389 (2017) 10.1073/pnas.1700141114 Hanashima (2007) Chem. Asian J. 2 1447 – 1459 (2007) 10.1002/asia.200600424 Hanashima (2007b) Org. Lett. 9 1777–1779 (2007) 10.1021/ol0704946 Hofmann (2015) Nature 526 241–244 (2015) 10.1038/nature15388 Kandasamy (2013) Chem. Commun. 49 4453-445 (2013) 10.1039/c3cc00042g Kandasamy (2014) Chem. Commun. 50 1875-1877 (2014) 10.1039/c3cc48860h Leonori (2013) Beilstein J. Org. Chem. 9 332–341 (2013) 10.3762/bjoc.9.38 Lisboa (2017) Proc .Natl. Acad. Sci. USA 114 11063-11068 (2017) 10.1073/pnas.1706875114 Martin (2013) J. Am. Chem. Soc. 135 9713−9722 (2013) 10.1021/ja401410y Martin (2013b) Chem. Commun. 49 7159-7161 (2013) 10.1039/c3cc43545h Nonaka (2014) Proc .Natl. Acad. Sci. USA 111 8173-8178 (2014) 10.1073/pnas.1319870111 Parameswarappa (2016) Cell Chem. Biol. 23 1407-1416 10.1016/j.chembiol.2016.09.016 Pereira (2015) Angew. Chem. Int. Ed. 54 10016 –10019 (2015) 10.1002/anie.201504847 Reinhardt (2015) Chem. Biol. 22 38–49 (2015) 10.1016/j.chembiol.2014.11.016 Schlegel (2011) ChemBioChem 12 2791 – 2800 (2011) 10.1002/cbic.201100511 Schmidt (2015) Chem. Eur. J. 21 5709 – 5713 (2015) 10.1002/chem.201500065 Schumann (2017) Sci. Transl. Med. Advance Online Publication 10.1126/scitranslmed.aaf5347 Seeberger (2017) Angew. Chem. Int. Ed. 56 13973 –13978 (2017) 10.1002/anie.201700964 Seeberger (2017b) Beilstein J. Org. Chem. 13 164–173 (2017) 10.3762/bjoc.13.19 Weishaupt (2013) Chem. Eur. J. 19 12497 – 12503 (2013) 10.1002/chem.201204518 Weishaupt (2016) Beilstein J. Org. Chem. 12 1440-1446 (2016) 10.3762/bjoc.12.139 Weishaupt (2017) Chem. Commun. Advance Online Publication 10.1039/C7CC00520B Yang (2012) Chem. Sci. 3 896-899 (2012) 10.1039/c1sc00804h Yang (2013) J. Am. Chem. Soc. 135 6262–6271 (2013) 10.1021/ja401164s
S39
Table S3: Additional published glycans in the library
Glycan ID
Glycan Structure (Microbial origin if applicable)
DOI Paper Saccharides
number
3
Man(a1-2)[Man(a1-6)Man(a1-6)]Man(a1-6)Man(a1-6)Man(a1-1)thiopentanol
(Mycobacterium tuberculosis)
10.1021/jo061233x 6
4
Araf(a1-5)Araf(a1-3)[Araf(a1-5)]Araf(a1-5)Araf(a1-5)Araf(a1-1)thiohexanol
(Mycobacterium tuberculosis)
10.1021/jo061233x 6
14
Fuc(a1-2)Gal(b1-3)GalNAc(b1-3)Gal(a1-4)Gal(b1-4)Glc(b1-1)thiopentanol
10.1021/ja069218x 6
16
GlcNAc(b1-1)thiopentanol
10.1002/cbic.201000020 1
17
Man(a1-2)Man(a1-1)thiopentanol
10.1002/cbic.201000020 2
19
Gal(b1-4)GlcNAc(b1-2)Man(a1-1)thiopentanol
10.1002/cbic.201000020 3
S40
21
Man(a1-6)Man(a1-4)GlcN(a1-6)Ino(1-P)phosphothiohexanol
(Plasmodium falciparum)
10.1038/nchembio.75 4
22
Man(a1-2)Man(a1-6)Man(a1-4)GlcN(a1-6)Ino(1-P)phosphothiohexanol
(Plasmodium falciparum)
10.1038/nchembio.75 5
23
Gal(b1-4)Glc(b1-1)thiopentanol
10.1002/cbic.201000020 2
24
Gal(a1-3)[Fuc(a1-2)]Gal(b1-4)GlcNAc(b1-1)thiopentanol
10.1002/cbic.201000020 4
25
Gal(b1-4)GlcNAc(b1-1)thiopentanol
10.1002/cbic.201000020 2
26
Gal(b1-2)Gal(b1-1)thiopentanol
10.1002/cbic.201000020 2
27
Gal(b1-3)Gal(b1-4)Glc(b1-1)thiopentanol
10.1002/cbic.201000020 3
28
Fuc(a1-2)Gal(b1-4)GlcNAc(b1-1)thiopentanol
10.1002/cbic.201000020 3
S41
29
Gal(b1-3)GlcNAc(b1-1)thiopentanol
10.1002/cbic.201000020 2
30
Gal-3-sulfate(b1-1)ThiolinkerA
10.1002/cbic.201000020 1
31
Gal-6-sulfate(b1-1-)ThiolinkerA
10.1002/cbic.201000020 1
32
GalNAc(b1-4)GlcNAc(b1-1)thiopentanol
10.1002/cbic.201000020 2
33
Gal(b1-3)GalNAc(a1-1)thiopentanol
10.1002/cbic.201000020 2
51
Gal(a1-1)PEG-Thiol
10.1021/bi8022703 1
52
Anthrose(b1-3)Rha(a1-3)Rha(a1-2)Rha(a1-1)thiopentanol
(Bacillus anthracis)
10.1111/j.1365-2672.2008.04129.x
4
53
Rha(a1-2)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
2
S42
54
Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
1
55
Anthrose(b1-3)Rha(a1-3)Rha(a1-1)thiopentanol
(Bacillus anthracis)
10.1111/j.1365-2672.2008.04129.x
4
56
Rha(a1-3)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
2
57
2-methyl-4-N-acetyl-4,6-dideoxyglucose(b1-3)Rha(a1-3)Rha(a1-2)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
4
58
Anthrose(b1-3)Rha(a1-1)thiopentanol
(Bacillus anthracis)
10.1111/j.1365-2672.2008.04129.x
2
S43
59
Anthrose(b1-1)thiopentanol
(Bacillus anthracis)
10.1111/j.1365-2672.2008.04129.x
1
60
2-Demethyl-Anthrose(b1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
1
61
2-Demethyl-Anthrose(b1-3)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
2
S44
62
2-Demethyl-Anthrose(b1-3)Rha(a1-3)Rha(a1-2)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
4
63
2-methyl-4-N-tert-butanoyl-4,6-dideoxyglucose(b1-3)Rha(a1-3)Rha(a1-2)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
4
64 2-methyl-4-N-isobutanoyl-4,6-dideoxyglucose(b1-3)Rha(a1-3)Rha(a1-2)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
4
S45
65
2-methyl-4-N-(3R-3-hydroxybutanoyl)l-4,6-dideoxyglucose(b1-3)Rha(a1-3)Rha(a1-2)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
4
S46
66
2-methyl-4-N-(3S-3-hydroxybutanoyl)l-4,6-dideoxyglucose(b1-3)Rha(a1-3)Rha(a1-2)Rha(a1-1)thiopentanol
10.1111/j.1365-2672.2008.04129.x
4
67
Man(a1-2)[Araf(a1-5)Araf(a1-3)[Araf(a1-5)]Araf(a1-5)Araf(a1-5)Araf(a1-2)[Man(a1-6)]Man(a1-6)]Man(a1-6)Man(a1-1)thiohexanol
l(Mycobacterium tuberculosis)
10.1021/jo061233x 13
68
Araf(a1-5)Araf(a1-1)thiohexanol
(Mycobacterium tuberculosis)
10.1021/jo061233x 2
S47
106
GlcN-6,N-disulfate(a1-4)IdoA-2-sulfate(a1-4)GlcN-6,N-disulfate(a1-4)IdoA-2-sulfate(a1-4)GlcN-6,N-disulfate(a1-4)IdoA-2-sulfate(a1-1)AminoLinker1
10.1002/chem.200601103 6
107
GlcNAc-6-sulfate(a1-4)IdoA-2-sulfate(a1-4)GlcNAc-6-sulfate(a1-4)IdoA-2-sulfate(a1-4)GlcNAc-6-sulfate(a1-4)IdoA-2-sulfate(a1-1)aminopentanol
10.1002/chem.200601103 6
108
GlcN-N-sulfate(a1-4)IdoA(a1-4)GlcN-N-sulfate(a1-4)IdoA(a1-4)GlcN-N-sulfate(a1-4)IdoA(a1-1)aminopentanol
10.1002/chem.200601103 6
109
GlcNAc(a1-4)IdoA(a1-4)GlcNAc(a1-4)IdoA(a1-4)GlcNAc(a1-4)IdoA(a1-1)aminopentanol
10.1002/chem.200601103 6
110
IdoA-2-sulfate(a1-4)GlcN-N-sulfate(a1-4)IdoA-2-sulfate(a1-4)GlcN-N-sulfate(a1-4)IdoA-2-sulfate(a1-4)GlcN-N-sulfate(a1-1)aminopentanol
10.1002/chem.200601103 6
111 IdoA-2-sulfate(a1-4)GlcNAc(a1-4)IdoA-2-sulfate(a1-4)GlcNAc(a1-4)IdoA-2-sulfate(a1-4)GlcNAc(a1-1)aminopentanol
10.1002/chem.200601103 6
S48
112
GlcN-6,N-disulfate(a1-4)IdoA-2-sulfate(a1-4)GlcN-6,N-disulfate(a1-4)IdoA-2-sulfate(a1-1)AminoLinker1
10.1002/chem.200601103 4
113
IdoA(a1-4)GlcN-N-sulfate(a1-4)IdoA(a1-4)GlcN-N-sulfate(a1-1)aminopentanol
10.1002/chem.200601103 4
114
IdoA(a1-4)GlcN(a1-4)GlcA(b1-3)GlcNAc(a1-1)AminoLinker1
10.1002/chem.200601103 4
115
GlcN-6,N-disulfate(a1-4)IdoA-2-sulfate(a1-1)AminoLinker1
10.1002/chem.200601103 2
116
IdoA-2,4-disulfate(a1-1)AminoLinker1
10.1002/chem.200601103 1
S49
117
GlcN-6,N-disulfate(a1-1)AminoLinker1
10.1002/chem.200601103 1
221
Glc(a1-4)GalNAc(b1-4)[Man-6-PEtN(a1-2)Man(a1-6)]Man(a1-4)GlcN(a1-6)Ino(1-P)phosphothiohexanol
(Toxoplasma gondii)
10.1002/anie.201406706 7
222
GalNAc(b1-4)[Man-6-PEtN(a1-2)Man(a1-6)]Man(a1-4)GlcN(a1-6)Ino(1-P)phosphothiohexanol
(Toxoplasma gondii)
10.1002/anie.201406706 6
223
GalNAc(b1-4)[Man-6-PEtN(a1-2)Man(a1-6)]Man-2-PEtN(a1-4)GlcN(a1-6)Ino(1-P)phosphothiohexanol
10.1002/anie.201406706 6
S50
224
GalNAc(b1-4)[Man(a1-2)Man(a1-6)]Man(a1-4)GlcN(a1-6)Ino(1-P)phosphothiohexanol
(Toxoplasma gondii)
10.1002/anie.201406706 6
228
GlcN(a1-6)Ino(1-P)phosphothiohexanol
10.1002/anie.201406706 2
259
Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P)phosphoaminopentanol
(Haemophilus influenzae)
10.1039/C7SC04521B 8
260
Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P)phosphoaminopentanol
(Haemophilus influenzae)
10.1039/C7SC04521B 12
261
Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P)phosphoaminopentanol
(Haemophilus influenzae)
10.1039/C7SC04521B 16
S51
262
Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P-3)Ribf(b1-1)ribitol(5-P)phosphoaminopentanol
(Haemophilus influenzae)
10.1039/C7SC04521B 20
264
Gal(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
10.1038/nature15388 3
265
Glc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
10.1038/nature15388 3
267
Araf(a1-5)Araf(a1-5)Araf(a1-6)Man(a1-6)Man(a1-6)Man(a1-1)aminopentanol
(Mycobacterium tuberculosis)
10.1073/pnas.1700141114 6
268
Gal(b1-3)[Rha(a1-4)]GalA(a1-2)Rha(a1-2)Rha(a1-2)Rha(a1-1)aminopentanol
(Klebsiella pneumoniae)
10.1002/anie.201700964 6
269
Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)GlcNAc(b1-1)aminopentanol
10.1016/j.chempr.2016.12.004 4
S52
270
Gal(b1-4)GlcNAc(b1-3)[Gal(b1-4)GlcNAc(b1-6)]Gal(b1-4)GlcNAc(b1-1)aminopentanol
10.1038/nature15388 6
271
Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)GlcNAc(b1-1)aminopentanol
10.1038/nature15388 6
272
Gal-6-sulfate(b1-4)GlcNAc(b1-3)Gal-6-sulfate(b1-4)GlcNAc(b1-1)aminopentanol
10.1016/j.chempr.2016.12.004 4
273
Gal-3,6-disulfate(b1-4)GlcNAc(b1-3)Gal-6-sulfate(b1-4)GlcNAc(b1-1)aminopentanol
10.1016/j.chempr.2016.12.004 4
274
Gal(b1-4)GlcNAc-6-sulfate(b1-3)Gal(b1-4)GlcNAc-6-sulfate(b1-1)aminopentanol
10.1016/j.chempr.2016.12.004 4
275
Gal-6-sulfate(b1-4)GlcNAc-6-sulfate(b1-3)Gal-6-sulfate(b1-4)GlcNAc-6-sulfate(b1-1)aminopentanol
10.1016/j.chempr.2016.12.004 4
S53
276
L-Araf(a1-3)[Xyl(b1-4)Xyl(b1-4)]Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)Xyl(b1-1)aminopentanol
10.1002/chem.201500065 7
277
L-Araf(a1-3)[L-Araf(a1-3)[Xyl(b1-4)]Xyl(b1-4)Xyl(b1-4)Xyl(b1-4)]Xyl(b1-4)Xyl(b1-1)aminopentanol
10.1002/chem.201500065 8
278
L-Araf(a1-3)[Xyl(b1-4)]Xyl(b1-4)Xyl(b1-1)aminopentanol
10.1002/chem.201500065 4
279
L-Araf(a1-3)Xyl(b1-4)Xyl(b1-4)Xyl(b1-1)aminopentanol
10.1002/chem.201500065 4
S54
280
Xyl(b1-2)L-Araf(a1-3)[Xyl(b1-4)]Xyl(b1-4)Xyl(b1-1)aminopentanol
10.1002/chem.201500065 5
281
L-Araf(a1-3)[L-Araf(a1-3)[Xyl(b1-4)Xyl(b1-4)]Xyl(b1-4)]Xyl(b1-4)Xyl(b1-1)aminopentanol
10.1002/chem.201500065 7
282
L-Araf(a1-3)[L-Araf(a1-3)[Xyl(b1-4)]Xyl(b1-4)Xyl(b1-4)]Xyl(b1-4)Xyl(b1-1)aminopentanol
10.1002/chem.201500065 7
S55
285
Gal(b1-3)[Rha(a1-4)]GalA(a1-1)aminopentanol
(Klebsiella pneumoniae)
10.1002/anie.201700964 3
286
Gal(b1-3)GalA(a1-1)aminopentanol
(Klebsiella pneumoniae)
10.1002/anie.201700964 2
287
Gal(b1-3)GalA(b1-1)aminopentanol
(Klebsiella pneumoniae)
10.1002/anie.201700964 2
288
Rha(a1-3)Gal(b1-1)aminopentanol
(Klebsiella pneumoniae)
10.1002/anie.201700964 2
289
Rha(a1-4)GalA(a1-1)aminopentanol
(Klebsiella pneumoniae)
10.1002/anie.201700964 2
290
Glc(b1-3)Rha(a1-3)Rha(a1-1)aminopentanol
(Streptococcus pneumoniae)
10.1021/jacs.7b07836 3
291
GlcA(a1-6)Glc(a1-2)Rha(a1-3)Rha(a1-1)aminopentanol
(Streptococcus pneumoniae)
10.1021/jacs.7b07836 4
S56
292
Rha(a1-3)Rha(a1-1)aminopentanol
10.1021/jacs.7b07836 2
293
Rha(b1-4)Glc(b1-3)Rha(a1-3)Rha(a1-1)aminopentanol
(Streptococcus pneumoniae)
10.1021/jacs.7b07836 4
294
Rha(b1-4)Glc(b1-1)aminopentanol
(Streptococcus pneumoniae)
10.1021/jacs.7b07836 2
295
GlcA(a1-6)Glc(a1-1)aminopentanol
(Streptococcus pneumoniae)
10.1021/jacs.7b07836 2
296
GlcA(a1-6)Glc(a1-2)[Rha(b1-4)Glc(b1-3)]Rha(a1-3)Rha(a1-1)aminopentanol
(Streptococcus pneumoniae)
10.1021/jacs.7b07836 6
297
Fuc(a1-2)Gal(b1-3)GalNAc(b1-3)Gal(a1-4)Gal(b1-4)Glc(b1-1)aminopentanol
10.1038/ncomms12482 6
299
Glc(a1-3)Glc(a1-3)Glc(a1-3)Glc(a1-6)Glc(a1-1)aminopentanol
10.1038/ncomms12482 5
S57
300
Glc(a1-4)Glc(a1-4)Glc(a1-4)Glc(a1-6)Glc(a1-1)aminopentanol
10.1038/ncomms12482 5
301
Glc(a1-6)Glc(a1-6)Glc(a1-6)Glc(a1-6)Glc(a1-1)aminopentanol
10.1038/ncomms12482 5
302
Glc(a1-3)Glc(a1-6)Glc(a1-6)Glc(a1-6)Glc(a1-1)aminopentanol
10.1038/ncomms12482 5
303
Glc(a1-6)Glc(a1-4)Glc(a1-4)Glc(a1-1)aminopentanol
10.1038/ncomms12482 4
304
Glc(a1-3)Glc(a1-3)Glc(a1-3)Glc(a1-1)aminopentanol
10.1038/ncomms12482 4
305
Gal(b1-3)GalNAc(b1-4)Gal(b1-4)Glc(b1-1)aminopentanol
10.1039/C5CC01368B 4
306
Gal(b1-3)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
10.1039/C5CC01368B 4
S58
307
Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
10.1039/C5CC01368B 4
308
Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
10.1039/C5CC01368B 6
309
Neu5Ac(a2-3)Gal(b1-3)GalNAc(b1-4)Gal(b1-4)Glc(b1-1)aminopentanol
10.1039/C5CC01368B 5
310
Neu5Ac(a2-3)Gal(b1-3)GlcNac(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
10.1039/C5CC01368B 5
311
Neu5Ac(a2-3)Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
10.1039/C5CC01368B 5
312
Neu5Ac(a2-3)Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)GlcNAc(b1-3)Gal(b1-4)Glc(b1-1)aminopentanol
10.1039/C5CC01368B 7
S59
314
LegA5Ac(b2-1)thioethanol
(Legionella pneumophila)
10.1021/jacs.5b00455 1
315
LegA5Ac7Ac(b2-1)thioethanol
(Legionella pneumophila)
10.1021/jacs.5b00455 1
320
GalA(a1-1)thiohexanol
10.1039/C3SC53362J 1
321
Gal-4,6-pyruvate(b1-1)thiohexanol
10.1039/C3SC53362J 1
322
Galf(b1-3)[AAT(a1-4)]GalNAc(a1-3)Gal-4,6-pyruvate(b1-1)thiohexanol
(Bacteroides fragilis)
10.1039/C3SC53362J 4
323
AAT(a1-4)GalA(a1-3)GalA(a1-1)thioethanol
(Streptococcus pneumoniae)
10.1039/C3SC53362J 3
S60
325
Glc(a1-4)Gal(a1-4)GlcA(b1-4)Glc(b1-4)Glc(a1-4)Gal(a1-1)aminoethanol
(Streptococcus pneumoniae)
10.1126/scitranslmed.aaf5347 6
326
Gal(a1-4)GlcA(b1-4)Glc(b1-4)Glc(a1-4)Gal(a1-1)aminoethanol
(Streptococcus pneumoniae)
10.1126/scitranslmed.aaf5347 5
327
Glc(b1-4)Glc(b1-4)Glc(a1-4)Gal(a1-1)aminoethanol
(Streptococcus pneumoniae)
10.1126/scitranslmed.aaf5347 4
328
Glc(b1-4)Glc(a1-4)Gal(a1-1)aminoethanol
(Streptococcus pneumoniae)
10.1126/scitranslmed.aaf5347 3
329
Glc(b1-4)Glc(b1-4)Glc(b1-1)aminopentanol
10.1039/C5OB02226F 3
330
Glc(b1-4)Glc(b1-4)Glc(b1-4)Glc(b1-4)Glc(b1-1)aminopentanol
10.1039/C5OB02226F 5
S61
331
Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)Glc(b1-1)aminopentanol
10.1039/C5OB02226F 4
332
Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)Glc(b1-1)aminopentanol
10.1039/C5OB02226F 6
333
Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)Glc(b1-1)aminopentanol
10.1039/C5OB02226F 8
S62
334
Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)Glc(b1-1)aminopentanol
10.1039/C5OB02226F 7
335
Xyl(a1-6)Glc(b1-4)[Xyl(a1-6)]Glc(b1-4)Glc(b1-4)Glc(b1-1)aminopentanol
10.1039/C5OB02226F 6
336
Gal(b1-3)Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 3
337
Gal(b1-3)Gal(b1-3)Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 4
338
Gal(b1-6)Gal(b1-6)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 3
S63
339
Gal(b1-3)[Gal(b1-6)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 4
340
Gal(b1-3)[Gal(b1-6)Gal(b1-6)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 5
341
Gal(b1-3)[Gal(b1-6)Gal(b1-6)Gal(b1-6)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 6
342
Gal(b1-3)Gal(b1-3)[Gal(b1-6)Gal(b1-6)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 6
343
L-Araf(a1-3)Gal(b1-3)Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 4
S64
344
Gal(b1-3)[L-Araf(a1-6)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 4
345
L-Araf(a1-3)[Gal(b1-6)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 4
346
Gal(b1-6)[L-Araf(a1-3)]Gal(b1-6)[Gal(b1-3)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 6
347
Gal(b1-6)[L-Araf(a1-3)]Gal(b1-6)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 4
S65
348
L-Araf(a1-3)Gal(b1-6)[Gal(b1-3)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 5
349
Gal(b1-6)[L-Araf(a1-3)]Gal(b1-6)Gal(b1-6)[Gal(b1-3)]Gal(b1-3)Gal(b1-1)aminopentanol
10.1021/acs.orglett.5b02185 7
350
Glc(b1-4)FucNAc(a1-3)D-FucNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
10.1073/pnas.1706875114 3
351
GlcA(b1-3)[Glc(b1-4)]FucNAc(a1-1)aminopentanol
(Streptococcus pneumoniae)
10.1073/pnas.1706875114 3
S66
352
GlcA(b1-3)[Glc(b1-4)]FucNAc(a1-3)D-FucNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
10.1073/pnas.1706875114 4
353
L-PneNAc(a1-2)GlcA(b1-3)[Glc(b1-4)]FucNAc(a1-3)D-FucNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
10.1073/pnas.1706875114 5
354
L-PneNAc(a1-2)GlcA(b1-3)[Glc(b1-4)]FucNAc(a1-3)QuiNAc(b1-1)aminopentanol
(Streptococcus pneumoniae)
10.1073/pnas.1706875114 5
S67
Table S4: Summary of linkers of glycans in the library
Thioethanol
Thiopentanol
Thiohexanol
Phosphothiohexanol
PEG-Thiol
ThiolinkerA
Aminoethanol
Aminopentanol
Aminohexanol
Aminododecanol
p-Aminocyclohexanol
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Aminolinker1
Aminolinker2
Thr/Threonine
Phosphoethanolamine
Phosphoaminopentanol
S69
Table S5: Estimated Kd values and avidity ranking for Burkholderia lectins towards aminopentanol-linked mannosides and heptosides
BC2L-A Rankd, BC2L-A BC2L-C-ct Rankd, BC2L-C-ct
Glycan Kd a Kd
Intensity
1 mg/mL
Intensity
0.4 mg/mL Kd
a Kd Intensity
1 mg/mL
Intensity
0.4 mg/mL
082 7.69 (0.280) ± 4.91 (0.178) 7 5 5 112.9 (4.53) ± 8.30 (0.33) 7 4 5
083 4.58 (0.167) ± 1.77 (0.0644) 3 4 4 97.2 (3.90) ±11.10 (0.45) 4 5 4
085 3.89 (0.141) ± 0.622 (0.0226) 2 2 2 64.53 (2.59) ± 15.59 (0.63) 1 3 2
226 5.90 (0.215) ± 2.66 (0.0970) 6 7 7 118.89 (4.77) ± 15.91 (0.64) 8 7 7
231 4.77 (0.173) ± 2.36 (0.0860) 4 6 6 110.02 (4.42) ± 13.46 (0.54) 6 6 6
237 3.66 (0.133) ± 1.30 (0.0474) 1 3 3 67.77 (2.72) ± 15.83 (0.64) 2 2 1
257 5.89 (0.214) ± 2.19 (0.0800) 5 1 1 82.47 (3.31) ± 18.58 (0.75) 3 1 3
092 n.d. b 12 12 >1050 (>42) c 14 13 14
093 n.d. 13 13 >1050 (>42) 15 16 16
094 382.9 (13.9) ± 112.6 (4.09) 11 11 11 301 (12.08) ± 116 (4.68) 11 11 11
095 n.d. 14 13 >1050 (>42) 13 14 13
096 n.d. 16 15 686 (27.54) ± 246 (9.9) 12 12 12
097 87.12 (3.17) ± 55.9 (2.03) 8 10 10 128 (5.12) ± 13.0 (0.52) 9 10 10
100 124.0 (4.51) ± 37.9 (1.38) 9 8 8 107 (4.31) ± 27 (1.1) 5 8 9
178 147.9 (5.48) ± 72.9 (2.65) 10 9 9 130 (5.22) ± 16.49 (0.66) 10 9 8
158 n.d. 15 16 >1050 (>42) 16 15 15
a Mean of three Kd values calculated from three glycan experiments. Format for Kd values: µg/mL (µM) ± SD from three experiments; molar Kd is based on the
concentration of the dimer with molecular weights of 27.5 kDa for BC2L-A and 24.9 kDa for BC2L-C-ct)..
b n.d. Signal intensities too low to determine dissociation constant (highest fluorescence intensities less than 5% of saturation intensity for 257)
c The value given for BC2L-C-ct is >1050 µg/mL (>42 µM) if not at least two data points of the concentration series were above the calculated Kd
d Ranking was performed from lowest to highest Kd values and from highest to lowest fluorescence intensities
S70
3. Supporting Figures
Figure S1: Cluster analysis of compounds of the MPS and the Imperial College London
Glycosciences Laboratory libraries (11). Each line terminus represents one glycan with MPS
glycans colored in orange, Imperial College glycans in blue, and structures contained in both
sets as well as inner branches leading to structures of both sets colored in green.
S71
Figure S2: Binding of eleven plant lectins to a glycan array printed with 140 synthetic
compounds. Values are the mean ± SD of four spots from two glycan array experiments
relative to the highest intensity glycan for each lectin. Glycans are arranged with expected
binders for each lectin to the left and color-coded according to SI Appendix Table S1 (see
Figure S3 for identical glycan arrangement for all lectins). Structures or class of strong
additional glycan ligands are indicated within the graphs. For several lectins, F1 represents
mono-D-FucNAc 102, F2 the mixture of mono-D-FucNAc with the oxidized form Sugp (glycan
195). For WGA, glycans marked with ‘a’ contain an internal GlcNAc residue that may mediate
binding (21). The complete information on which glycan was bound by which lectin can be
found in the list of glycans (SI Appendix Table S2).
S72
Figure S3: Comparison of lectin binding profiles for eleven plant lectins to a glycan array printed
with 140 synthetic compounds. Values are the mean ± SD of four spots from two glycan array
experiments relative to the highest intensity glycan for each lectin. Glycans are arranged
identically in each panel, see Figure S2 for a comparison between observed and predicted
binding. All intensity values are listed in the Supporting Data files.
S73
Figure S4: Binding of antibodies from the sera of 15 healthy individuals to the complete set of 140 immobilized glycans. Data are presented as
boxplots with each value being the mean of four spots from two glycan experiments normalized to the mean binding signal of each experiment. The
black bar represents the median, the red bar the mean fluorescence intensity of the 15 sera.
S74
Figure S5: Binding of Fc-Fusion protein DC-SIGN-D to selected compounds on the 140
compound glycan array. All signals are normalized to the intensity of LeY tetrasaccharide 157
at a protein concentration of 25 µg/mL. Error bars represent SD of four spots from two glycan
array experiments.
S75
Figure S6: Complete binding profile of DC-SIGN-T on 140 glycan array at 25 µg/mL. Each
value is the mean of four spots from two glycan array experiments normalized to the intensity
of LeY tetrasaccharide 157 at 25 µg/mL. Only structures of glycans that are not included in
main text Figure 5 are shown. Tetrasaccharide 250 is identical to 82 except that it bears an
aminoethanol instead of an aminopentanol linker.
S76
Figure S7: Complete binding profile of DC-SIGN-T on 140 glycan array at 5 µg/mL. Each value
is the mean of four spots from two glycan array experiments normalized to the intensity of LeY
tetrasaccharide 157 at 25 µg/mL.
S77
Figure S8: Complete binding profile of DC-SIGN-T on 140 glycan array at 1 µg/mL. Each value
is the mean of four spots from two glycan array experiments normalized to the intensity of LeY
tetrasaccharide 157 at 25 µg/mL.
S78
Figure S9: Complete binding profile of DC-SIGN-D on 140 glycan array at 25 µg/mL. Each
value is the mean of four spots from two glycan array experiments normalized to the intensity
of LeY tetrasaccharide 157 at 25 µg/mL. Only structures of glycans that are not included in SI
Appendix Figure S3 are shown. Tetrasaccharide 250 is identical to 82 except that it bears an
aminoethanol instead of an aminopentanol linker.
S79
Figure S10: Complete binding profile of DC-SIGN-D on 140 glycan array at 5 µg/mL. Each
value is the mean of four spots from two glycan array experiments normalized to the intensity
of LeY tetrasaccharide 157 at 25 µg/mL.
S80
Figure S11: Complete binding profile of DC-SIGN-D on 140 glycan array at 1 µg/mL. Each
value is the mean of four spots from two glycan array experiments normalized to the intensity
of LeY tetrasaccharide 157 at 25 µg/mL.
S81
Figure S12: Interaction of DC-SIGN-T with immobilized LPS inner core diheptoside 178.
Sensorgrams (a) and the resulting saturation plot (b) are shown. (c) Resulting apparent (app)
Kd and Rmax values are indicated. Kd is the mean ± SD of two independent measurements.
S82
Figure S13: Binding of BC2L-A and BC2L-C-ct at 1 mg/mL to the 140 compound glycan array. Each value in the barchart is the mean ± SD of six
spots from three glycan experiments with the insets showing representative field images. Terminal mannosides and heptosides are labeled with M
and H, respectively. M* denotes terminal mannosides in which this saccharide is modified with a phosphoethanolamine.
S83
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