Methods for generation of monoclonal antibodies to the very small drug hapten, 5-benzimidazolecarboxylic acid.
Drug-specific monoclonal antibodies (MAbs) were produced against the very small drug hapten (162.15 Da), 5-benzimidazolecarboxylic acid, an analogue of 2-(4-Thiazolyl)benzimidazole (TBZ) but lacking the thiol group. TBZ is widely used as a broad-spectrum anthelmintic in various animal species and humans and also as a food preservative and agricultural fungicide. The anti-5-benzimidazolecarboxylic acid antibodies produced have potential use for extraction and/or detection of protein-bound residue forms of TBZ. Three in vivo immunisation regimes (with combinations of two related small drug haptens and two different adjuvants/carrier molecules) and an in vitro immunisation procedure using a combination of three related unconjugated small drug haptens were investigated. Specificity for the hapten immunogen/s was initially determined using two different ELISA procedures. BIACORE analysis, in conjunction with drug binding inhibition studies, was used to confirm the specificity of a small number of selected clones. In vivo immunisation with a drug molecule conjugated to a lipopeptide/T-cell epitope, which acts both as a carrier molecule and an adjuvant was the most useful of the methods tested for the production of specific MAbs to a typically very small hapten with low immunogenic properties.
Moran
E
E
National Cell and Tissue Culture Centre, Dublin City University, Glasnevin, Ireland. moranl@biores-irl.ie
O'Keeffe
M
M
O'Connor
R
R
Larkin
A M
AM
Murphy
P
P
Clynes
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000937:Antigen-Antibody Reactions / Q000276:immunology; D001562:Benzimidazoles / Q000276:immunology*; D002264:Carboxylic Acids / Q000276:immunology*; D002352:Carrier Proteins / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000276:immunology*; D007114:Immunization / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D020349:Surface Plasmon Resonance
C470231:5-benzimidazolecarboxylic acid; D000911:Antibodies, Monoclonal; D001562:Benzimidazoles; D002264:Carboxylic Acids; D002352:Carrier Proteins; D006241:Haptens