Abstract
Our experience in coating of solid surfaces with glycans, mainly for obtaining routine glycoarrays based on immunological plates, is summarized. Three polystyrene coating techniques are described: direct physical adsorption, covalent binding, and immobilization using the biotin tag. Protocols for studies on anticarbohydrate antibodies are considered, with special emphasis on the application niches of different immobilization techniques as related to the specificity of each method of glycan-binding protein assay, as well as the problems of background binding and quantitative estimation of the results.
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Abbreviations
- Atri :
-
A trisaccharide, GalNAcα1-3(Fucα1-2)Galβ-
- AP:
-
Alkaline phosphatase
- Btri :
-
B trisaccharide, Galα1-3(Fucα1-2)Gal-
- Biot:
-
Biotin
- BSA:
-
Bovine serum albumin
- ELISA:
-
Enzyme-linked immunosorbent assay
- Glyc:
-
Glycoside residue
- GPC:
-
Gel-permeation
- Ig:
-
Anti-mouse antibodies
- HRPO:
-
Horse radish peroxidase
- m.w.:
-
Molecular weight
- PAA:
-
Poly(acrylamide)
- PBS:
-
Phosphate saline buffer
- pNPA:
-
poly(4-nitrophenylacrylate)
- pNSA:
-
poly(N-oxisuccinimidylacrylate)
- Str:
-
Streptavidin
- TLC:
-
Thin-layer chromatography
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Acknowledgments
This study was supported by the Program of the Presidium of the RAS “Molecular and Cell Biology” (FP6 Project SP5B-CT-2007-044512) and the RFBR grant #10-04-01693-a.
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Galanina, O.E., Chinarev, A.A., Shilova, N.V., Sablina, M.A., Bovin, N.V. (2012). Immobilization of Polyacrylamide-Based Glycoconjugates on Solid Phase in Immunosorbent Assays. In: Chevolot, Y. (eds) Carbohydrate Microarrays. Methods in Molecular Biology, vol 808. Humana Press. https://doi.org/10.1007/978-1-61779-373-8_12
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DOI: https://doi.org/10.1007/978-1-61779-373-8_12
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