University of Surrey

Test tubes in the lab Research in the ATI Dance Research

The measurement of a glycated protein by immunoassay.

Kirkham, Paul A. (1989) The measurement of a glycated protein by immunoassay. Doctoral thesis, University of Surrey (United Kingdom)..

[img]
Preview
Text
10800223.pdf
Available under License Creative Commons Attribution Non-commercial Share Alike.

Download (10MB) | Preview

Abstract

The measurement of glycated proteins, in particular, glycated albumin using specific antibodies offers the much-needed possibility of a (semi) automated method for the diagnosis and monitoring of diabetic control. Glycated proteins would appear to be poor immunogens as is evident by the lack of antibodies successfully raised against them when compared against the number of antibodies recognising reduced-glycated proteins. This study has successfully overcome this problem by synthesing several different glycated compounds as haptens and then using them in conjunction with a carrier protein to raise antisera in sheep. The binding of one of these antisera, with a titre 1:100000 showed considerable displacement when incubated with diabetic plasma at various dilutions. Western blot analysis on human plasma confirmed that the antiserum specifically recognised a continuous epitope on glycated human serum albumin. Affinity purified antibodies were used to develop both an indirect competitive ELISA and later a direct non-competitive ELISA for glycated serum albumin which does not require prior reduction of the glycated protein to the glucitol form. These assays have a dynamic range at 0 to 100 mug/ml and 0 to 50mug/ml of glycated human serum albumin respectively. The competitive ELISA exhibited <0.15% cross reactivity with both sodium borohydride and sodium periodate treated human serum albumin. Further work was needed to be undertaken to develop a rugged ELISA that could discriminate between diabetics and normals by routinely measuring glycated human serum albumin levels.

Item Type: Thesis (Doctoral)
Divisions : Theses
Authors :
NameEmailORCID
Kirkham, Paul A.
Date : 1989
Contributors :
ContributionNameEmailORCID
http://www.loc.gov/loc.terms/relators/THS
Additional Information : Thesis (Ph.D.)--University of Surrey (United Kingdom), 1989.
Depositing User : EPrints Services
Date Deposited : 22 Jun 2018 13:56
Last Modified : 06 Nov 2018 16:53
URI: http://epubs.surrey.ac.uk/id/eprint/847606

Actions (login required)

View Item View Item

Downloads

Downloads per month over past year


Information about this web site

© The University of Surrey, Guildford, Surrey, GU2 7XH, United Kingdom.
+44 (0)1483 300800