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Methods Suitable for Large Scale Quantitation of Sugar Residues in Microheterogenic Glycoproteins
Cervén, Erik
Cervén, Erik
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Issue Date
1991
Submitted date
2024-02-28
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Abstract
Three methods suitable for the purpose of large scale quantitation of sugar residues in samples of microheterogenic glycoproteins are presented, named by the generic names 1) Marked-Lectin Immunoassay (MALIA), 2) Oxidative-Reductive Immunoassay (Redox-IA), or 3) relying on incorporation of labeled sugars from precursors using serum glycosyltransferases. All three methods can be used in the presence of serum and comprise only simple incubations and washings that would be possible to perform with standard or slightly modified RIA or ELISA equipment. 1) In MALIA, the glycoproteins are bound to an immunoadsorbentandthis is used in combination with an incubation with marked lectin. The utility of the method is restricted only by the specificity of the lectin and its affinity for the solid phase antigen during washing. When thelectin has low affinity for the sugar residues, they can be measured in the washing supernatants with a similar sensitivity of method. 2) In Redox-IA, the contentofsialic acid in a glycoprotein bound from a body fluid onto an immunoadsorbent is measured using the increased Susceptibility of sialic acid residues to oxidation which are a) oxidized with prefered specificity and b) reduced using a labeled reductant followed by c) isolation of the labeled species. The utility of the method is restricted to measurementsof averagesialic acid content in glycoproteins whose identity is specified by their affinity to the immunoadsorbent. 3) Using the endogenous serum glycosyltransferases and labeled nucleotide-linked sugars, crude information can be obtained about the stage of glycosylation of the serum components. The previously predominant method to precipitate the labeled species with acid has been replaced by a method using solid phase lectins onto which the labeled product is collected with preserved immunospecificity.
Citation
Protein glycosylation, 189 - 192
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Book chapter
conference paper
conference paper
Language
en
Description
Series/Report no.
GBF monographs ; Volume 15
ISSN
0930-4320
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ISBN
1560811846
3527283676
3527283676
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Attribution-NonCommercial-ShareAlike 4.0 International