Production of Polyclonal and Monoclonal Antibodies for Detection of Sulfonamides.

摘要

Polyclonal antibodies were produced in rabbits by immunizing them with sulfamethazine conjugated via a glutoraldehyde bridge to bovine serum albumin. These antibodies were semi-purified by precipitation with ammonium sulfate. Cross-reactivity studies reveal that the antibodies are highly specific for sulfamethazine. They do not show any cross-reactivity with any closely related drugs or with chlortetracycline and penicillin, which are added, in combination with sulfamethazine, to swine feed. These antibodies offer excellent sensitivity in the direct competitive ELISA. As little as 5 parts per billion sulfamethazine can be detected. A twenty minute laboratory assay was developed for use as a quick screening qualitative assay or to quantitate sulfamethazine residues in swine urine. A study, performed on the enzyme conjugate, indicates that it will offer long-term stability. A mouse immunized intraperitoneally with sulfamethazine-bovine serum albumin conjugate was used as the spleen cell donor for fusion. Hybridoma cell lines which secrete anti-sulfamethazine antibody are being identified and evaluated. Commercial applications of the research include: (1) laboratory assays for detection of sulfamethazine in milk and tissue, and (2) a kit for quick screening for field use.