Comparison of carbohydrate and peptide biotinylation on the immunological activity of IgG1 murine monoclonal antibodies.
When the classical amino acid esterification procedure was used for the biotinylation of the IgG1 monoclonal antibody J28 it resulted in a loss of immunological activity. This antibody recognizes the fetoacinar pancreatic (FAP) antigen and the decrease in reactivity was directly proportional to the molar biotin/antibody ratio indicating substitutions at or near the antibody combining site. This effect was specific to J28 since the IgG1 Mab F22 which recognises the same antigen was not damaged by this procedure. Active Mab J28 conjugates were obtained using biotinylation via oligosaccharide moieties. The biotinylation efficiency using this method was dependent on the previous degree of antibody periodate oxidation and the maximal substitution was 3 mol biotin per mol of antibody. Using these conditions the sensitivity of the biotinylated J28 for the FAP antigen was similar to that obtained when using non-substituted antibody in the two antibodies technique.
Miralles
F
F
Laboratoire d'Immunochimie, Institut de Recherches Scientifiques sur le Cancer, C.N.R.S., Villejuif, France.
Takeda
Y
Y
Escribano
M J
MJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology; D001710:Biotin / Q000737:chemistry*; D002352:Carrier Proteins / Q000032:analysis / Q000276:immunology; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology; D008049:Lipase; D051379:Mice; D009844:Oligosaccharides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D002352:Carrier Proteins; D006023:Glycoproteins; D007074:Immunoglobulin G; D009844:Oligosaccharides; D010455:Peptides; C031192:oncofetal antigens; D001710:Biotin; C054621:CEL protein, human; D008049:Lipase