PMID: 2493509Feb 24, 1989Paper

Solid-phase immunoassay of serum antibodies to peptides. Covalent antigen binding to adsorbed phenylalanine-lysine copolymers

Journal of Immunological Methods
R N Hobbs

Abstract

Two small peptide antigens, glucagon and enkephalin (5-L-leucine), were covalently immobilised using either glutaraldehyde or bis-(sulphosuccinimidyl) suberate to an adsorbed layer of phenylalanine-lysine copolymer (PL) or partially acetylated PL (APL) on polystyrene. Both copolymers formed stable layers, particularly APL after adsorption from solution in distilled water. Adsorption of the copolymers under these conditions and subsequent coupling of the antigens yielded solid phases with low non-specific immunoglobulin binding characteristics in an enzyme-linked immunosorbent assay (ELISA) to detect peptide-specific antibodies in rabbit serum. The signal-to-noise ratio in this ELISA was dependent on the combination of copolymer, antigen and coupling reagent employed. Removal from the solid-phase of weakly bound antigen by washing with solutions containing Tween 20 or sodium dodecyl sulphate (SDS) increased assay sensitivity, which was 2-4-fold greater than when simple antigen adsorption was utilised. In the ELISA, the coefficient of variation was lower when covalent antigen coupling was employed.

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Citations

Apr 1, 1999·Journal of Peptide Science : an Official Publication of the European Peptide Society·K GregoriusH I Elsner
Jul 26, 1991·Journal of Immunological Methods·P SithigorngulC Cowden
Feb 26, 1993·Journal of Immunological Methods·A NiveleauR Dante
Feb 3, 1993·Journal of Immunological Methods·S C SmithK James
Apr 12, 1995·Journal of Immunological Methods·K GregoriusH I Elsner
May 2, 1994·Journal of Immunological Methods·J M BallM J Newman
Jan 1, 1991·Journal of Neuroscience Methods·W S ChangN T Neff
Jan 1, 1989·Current Opinion in Immunology·B Nilsson
Oct 16, 2002·Journal of Immunological Methods·José Vidal
Aug 10, 2000·Journal of Immunoassay·J E Butler
Nov 24, 2004·Journal of Immunoassay & Immunochemistry·José Vidal
Oct 8, 1999·Journal of Chromatography. B, Biomedical Sciences and Applications·J YangJ Xiang

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