Preparation Of Clenbuterol Monoclonal Antibody And Development Of Immunochromatographic Strips For CL Test

Clenbuterol hydrochloride is a sympathomimetic beta2- adrenergic agonist, also named "leanness enhancer", "repartitioning agent" in animal industry. It's widely used for the treatment of asthma in both humans and animals. On the other hand, it has a powerful effect of promoting animal growth with increased accretion of skeletal muscle mass and decreased accretion of body fat. For this reason, it was widely used as feed additive in animal production. Accumulative residue of clenbuterol in animal products can cause symptoms of acute poisoning in people, therefore its addition to animal feeds has been forbidden in many countries for security reason.Illegal use of CL is mainly detected by chromatography and immunoassay at present. Both methods are complicated, inconvenient, costly, and need professional skills, which causes detection can not be performed in shambles. In this study, three clenbuterol monoclonal antibodies are prepared by cell fusion technology, and test strips of immunochromatographicy are developed. Thereby, detection of CL becomes fast, simple and convenient.CL is coupled with Ovalbumin (OVA) or Bovine Serum Albumin (BSA) by diazotization, producing the immune antigen (BSA-CL) and the coating antigen (OVA-CL). They are identified with UV scanning and SDS-PAGE. In the results, absorbent wavelength of carrier proteins (OVA, BSA) has been changed after conjugation, and we can know from electrophoresis that molecular weight of conjugant is larger than that of carrier proteins. From these, we can know CL has been conjugated to carrier proteins.Immunization of Balb/C mice with the conjugant is carried out by hypodermic injection. Antisera of mice are detected by indirect enzyme-linked immunosorbent assay (ELISA) with...