In situ Modification of Immobilized Carbohydrates

Beside qualitative detection of oligosaccharides and quantitative affinity or kinetic analyses, the in situ modification of carbohydrates immobilized at a sensor chip surface has been published [6, 19], Van der Merwe et al. [20] used this approach to analyze the binding of B-lymphocyte antigen, CD22, to the highly glycosylated leukocyte surface protein CD45. 

Native CD45 (CD45-thy) purified from rat thymus was covalently coupled to sensor chip CMS. CD22Fc (domains 1-3 of mouse CD22 fused to the Fc portion of human IgG) interacted with immobilized CD45-thy. The in situ modification of the sialoglycoconjugates present on CD45-thy by desialylation and resialylation provided the basis for a detailed characterization of this interaction. 

Jonsson U. and Malmqvist M. (1992) Real time biospecific interaction analysis. The integration of surface plasmon resonance. Detection, general biospecific interface chemistry and microfluidics into one analytical system, Adv. Biosensors 2 291-336. 

Karlsson R. and Stahlberg R. (1995) Surface plasmon resonance detection and multi-sensing for direct monitoring of interactions involving low molecular weight analytes and for determination of low affinities. Anal. Biochem. 228 274 280. 

Karlsson R. and Fait A. (1997) Experimental design for kinetic analysis of protein protein interactions with surface plasmon resonance biosensors, J. Immunol. Methods 200 121-133. 

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