Journal: Polysaccharides

Article Title: Carbohydrate Microarrays

doi: 10.1007/978-3-319-16298-0_35

Figure Lengend Snippet: A high-throughput platform for carbohydrate-based microarrays. A high-precision robot designed to produce cDNA microarrays is utilized to spot carbohydrate antigens of various structural configurations onto a nitrocellulose-coated glass slide. The microspotting capacity of this system is approximately 20,000 spots per chip. The antibody- or GBP-stained slides were then scanned for fluorescent signals with a microarray scanner that was developed for cDNA microarrays. Toxins or viral pathogens can also be applied on this platform of carbohydrate microarrays to probe potential glycan-receptors of viruses or toxins

Article Snippet: These include polysaccharide and glycoconjugate microarrays, reported by Denong Wang’s group at Columbia University’s Genome Center (now at SRI International, CA, USA) (Kiessling and Cairo ; Wang et al. ); monosaccharide chips, by Milan Mrksich and coworkers at the University of Chicago (Houseman and Mrksich ); arrays of natural and synthetic neoglycolipids, by Ten Feizi’s group at Imperial College Faculty of Medicine, Harrow, UK (Fukui et al. ); and arrays of synthetic oligosaccharides in microtiter plates, by a Scripps Research Institute group led by Chi-Huey Wong (Bryan et al. ).

Techniques: High Throughput Screening Assay, Staining, Microarray, Glycoproteomics

Journal: Polysaccharides

Article Title: Carbohydrate Microarrays

doi: 10.1007/978-3-319-16298-0_35

Figure Lengend Snippet: A carbohydrate microarray analysis of oligomannosyl antigens for expression of the 2G12-like and GNA-like glyco-epitopes. The mannose-cluster-containing microarrays were stained with 2G12 (5 μg/ml) and a biotinylated GNA (1.0 μg/ml), respectively. The former was revealed by Alexa 647 -tagged Goat anti-human IgG Fc-specific antibodies at 2 μg/ml and then developed with Streptavidin-Cy5 conjugate at 2 μg/ml. ( a ) Shows microarray images stained with either GNA or 2G12. ( b ) Illustrates the cluster configuration of the three Man9-conjugates. Microarray detection signals are shown as the mean fluorescent intensities (MFIs) of each microspot as captured by the ScanArray 5000A for the arrays stained with GNA ( c ) and 2G12 ( d ), respectively. Results were compared using overlay plots of the MFIs of staining signal ( red circles ) versus those of local backgrounds surrounding the antigen microarrays ( blue +) ( c , d ) (Adapted from Wang et al. )

Article Snippet: These include polysaccharide and glycoconjugate microarrays, reported by Denong Wang’s group at Columbia University’s Genome Center (now at SRI International, CA, USA) (Kiessling and Cairo ; Wang et al. ); monosaccharide chips, by Milan Mrksich and coworkers at the University of Chicago (Houseman and Mrksich ); arrays of natural and synthetic neoglycolipids, by Ten Feizi’s group at Imperial College Faculty of Medicine, Harrow, UK (Fukui et al. ); and arrays of synthetic oligosaccharides in microtiter plates, by a Scripps Research Institute group led by Chi-Huey Wong (Bryan et al. ).

Techniques: Microarray, Expressing, Staining

Journal: Polysaccharides

Article Title: Carbohydrate Microarrays

doi: 10.1007/978-3-319-16298-0_35

Figure Lengend Snippet: Integrated lipid/carbohydrate arrays recognize globally elevated antigen-specific antibodies in the CSF of MS patients as compared to those detected in the OND CSF. CSF samples from 11 MS (10 RRMS and 1 SPMS) and 9 OND subjects were characterized using microarrays spotted with 12 lipid and 20 carbohydrate antigens. Each preparation was spotted in triplicate at 2–4 dilutions. As illustrated in the overlay plots ( a – d ), this microarray supports detection of 126 unique antibody signatures. These included 63 IgG ( a , c ) and 63 IgM ( b , d ) signatures. Anti-human IgG or IgM secondary antibodies were used to reveal the antigen-specific antibodies detected by these microarrays. The captured IgG were stained with an anti-human IgG antibody conjugated with Cy3 at 2 μg/ml and the captured IgM in the same array revealed by a biotinylated anti-human IgM secondary antibody at 2 μg/ml and developed with Streptavidin-Cy5 conjugate at 2 μg/ml. Microarray datasets in ( a ) and ( b ) were illustrated as microarray scores. In ( c ) and ( d ) microarray datasets were further processed using JMP Genomics to produce RAR* scores. The results are presented as overlay plots of the mean microarray scores for each group. The colored needles that link the group mean values of each pair of scores provide a global comparison of the antibody profiles of MS ( red circles ) and OND ( blue crosses ) groups (Adapted from Wang et al. ). * RAR Score : RAR R elative A ntibody R eactivity, the value of log2 transformed and IQR-standardized microarray value (mean-background). For each antigen in a given concentration, the mean value of triplicate array detections was calculated

Article Snippet: These include polysaccharide and glycoconjugate microarrays, reported by Denong Wang’s group at Columbia University’s Genome Center (now at SRI International, CA, USA) (Kiessling and Cairo ; Wang et al. ); monosaccharide chips, by Milan Mrksich and coworkers at the University of Chicago (Houseman and Mrksich ); arrays of natural and synthetic neoglycolipids, by Ten Feizi’s group at Imperial College Faculty of Medicine, Harrow, UK (Fukui et al. ); and arrays of synthetic oligosaccharides in microtiter plates, by a Scripps Research Institute group led by Chi-Huey Wong (Bryan et al. ).

Techniques: Microarray, Staining, Comparison, Transformation Assay, Concentration Assay