Directional coupling of synthetic peptides to poly-L-lysine and applications to the ELISA.
Synthetic peptides are increasingly being used as antigens to generate highly specific antisera. Screening the antipeptide sera by enzyme-linked immunosorbent assay (ELISA) can suffer from carrier crossreactivity or lack sensitivity due to poor adsorption or presentation of the peptides. In this work we describe a procedure utilizing a heterobifunctional crosslinker to effect the directional coupling of synthetic peptides to poly-L-lysine preadsorbed to microtiter plates. Plates prepared by this method conferred precision, sensitivity, and specificity to an ELISA for antipeptide antisera. In a competitive ELISA format this method permitted detection of specific peptides to 3.7 x 10(-10) M and provided an assay sensitive to protein structure in solution.
Department of Biochemistry and Biophysics, University of California, Davis 95616.
C V
CV
Gegg
M E
ME
Etzler
D000595:Amino Acid Sequence; D000818:Animals; D003545:Cysteine / Q000737:chemistry; D004355:Drug Stability; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D008297:Male; D008832:Microchemistry / Q000379:methods; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000737:chemistry; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry*; D011817:Rabbits; D012680:Sensitivity and Specificity
D010455:Peptides; D011107:Polylysine; D010047:Ovalbumin; D003545:Cysteine