Measurement of antibody binding to antigenic peptides conjugated in situ to albumin-coated microtitre plates.
Monoclonal antibodies have been prepared against a synthetic peptide with a sequence corresponding to a repeated hydrophilic region of the protein core of the human MUC-2 gastrointestinal mucin. Peptide conjugates, prepared by glutaraldehyde cross-linking with keyhole limpet haemocyanin (KLH) and bovine serum albumin (BSA), were employed as the immunogen and target antigen (for screening by ELISA), respectively. However, for the measurement of antibody binding to peptide by an ELISA procedure, an alternative strategy was developed and is described in this report: peptides were conjugated directly to BSA immobilized by physical adsorption to the surface of microtitre plate wells. This procedure permits peptides to be tested as target antigens by ELISA without prior preparation of peptide-carrier conjugates.
Price
M R
MR
Cancer Research Laboratories, University of Nottingham, UK.
Sekowski
M
M
Hooi
D S
DS
Durrant
L G
LG
Hudecz
F
F
Tendler
S J
SJ
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D055262:Mucin-2; D009077:Mucins / Q000276:immunology; D009363:Neoplasm Proteins / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000276:immunology*
D000911:Antibodies, Monoclonal; C524625:MUC2 protein, human; D055262:Mucin-2; D009077:Mucins; D009363:Neoplasm Proteins; D012710:Serum Albumin, Bovine