Fast immunopurification of small amounts of specific antibodies on peptides bound to ELISA plates》.pdf

Journal oflmmunological Methods, 162 (1993) 69-75 69 © 1993Elsevier Science Publishers B.V. All rights reserved 0022-1759/93/$06.00 JIM 06698 Fast immunopurification of small amounts of specific antibodies on peptides bound to ELISA plates Karima Brahimi a, Jean-Louis P6rignon a, Marc Bossus b, H61~ne Gras b, Andr6 Tartar b and Pierre Druilhe a a Parasitologie Bio-M~dicale, Institut Pasteur, Paris, France, and b Laboratoire de Chimie des Biomol~cules, Institut Pasteur, Lille, France (Received 4 January 1993,accepted 1 February 1993) ELISA is widely used as a means to detect antibodies, but the potential of ELISA plates as an immunosorbent for the purification of specific antibodies does not seem to have been evaluated. In this study, ELISA plates coated with peptides representing short sequences of various antigens from Plasmodium falciparurn, the etiologic agent of human malaria, have been successfully used as a means to purify small amounts of the corresponding antibodies. ELISA plates, identical to those used for antibody detection, also permitted the evaluation of various elution conditions for each pairing of peptide and serum; we tested four eluting buffers (0.2 M glycine, pH 2.5; 0.2 M lysine, pH 11.5; 3.0 M MgCI2, 0.075 M Hepes, 25% ethylene glycol, pH 7.1-7.2 and 4 M NHaSCN in 0.1 M NaHzPO4, pH 6.0) with four pairs of peptides and sera. The ELISA plates could also be used to est