Haptenated nylon-coated polystyrene plates as a solid phase for ELISA.
An ELISA system, based on the novel use of a hapten-nylon conjugate as solid-phase coating antigen, has been applied in the screening of hybridoma cultures for anti-hapten monoclonal antibodies directed against the herbicide atrazine and its derivatives. Conjugation of a 2-aminocaproic acid derivative of atrazine with DCC to polyamide (Nylon 6) gave haptenated nylon which was soluble in aqueous cresol-ethanol mixtures and adsorbed efficiently on polystyrene microtitre plates. Reproducible ELISA results were obtained with culture supernatants of hybridomas derived from spleen cells of mice that had been immunized with atrazine-bovine serum albumin conjugates. Satisfactory results were also obtained with a water soluble peptide conjugated to nylon for use as a coating antigen in an ELISA. Plates coated with hapten-nylon as antigen have the added advantage that they can be stored at room temperature for at least 6 months without loss of activity. Nylon therefore appears to have general applicability as a carrier for both non-polar and polar haptens in the preparation and use of coating antigens.
Verschoor
J A
JA
Department of Biochemistry, University of Pretoria, Republic of South Africa.
Vermeulen
N M
NM
Visser
L
L
D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D004024:Dicyclohexylcarbodiimide; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000276:immunology*; D007075:Immunoglobulin M / Q000032:analysis; D051379:Mice; D009757:Nylons; D011137:Polystyrenes
D000911:Antibodies, Monoclonal; D006241:Haptens; D007075:Immunoglobulin M; D009757:Nylons; D011137:Polystyrenes; D004024:Dicyclohexylcarbodiimide