title: Production of monoclonal antibody against clonazepam for immunoassay of benzodiazepine drugs in swine tissues.
abstract: The objective of the present study was to produce a generic monoclonal antibody for immunoassay of residues of benzodiazepine drugs in swine tissues. Clonazepam was used to synthesize a hapten that was coupled to bovine serum albumin as an immunogen for the production of monoclonal antibody. Results showed that the obtained monoclonal antibody was able to recognize five benzodiazepine drugs simultaneously (clonazepam, flunitrazepam nitrazepam, diazepam, and oxazepam). The cross-reactivities were in the range of 24-100% and the limits of detection were in the range of 0.2-1.5 ng mL(-1) depending on the drug. Then a competitive indirect enzyme-linked immunosorbent assay was developed to determine the residues of five benzodiazepines in swine tissues (muscle, liver and kidney). The recoveries of five analytes from the fortified blank samples were in the range of 74.5-96.5% with coefficients of variation lower than 16.7%. Therefore, this immunoassay could be used as a rapid and simple method for the screening of residues of five benzodiazepine drugs in animal-derived foods.
authors.affiliation: a College of Veterinary Medicine , Agricultural University of Hebei , Baoding , Hebei , China.
authors.forename: Wen C
authors.initials: WC
authors.lastname: Shan
authors.forename: Ya L
authors.initials: YL
authors.lastname: Cui
authors.forename: Xin
authors.initials: X
authors.lastname: He
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Jian P
authors.initials: JP
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001569:Benzodiazepines / Q000032:analysis*; D002998:Clonazepam / Q000032:analysis; D003975:Diazepam / Q000032:analysis; D004350:Drug Residues / Q000032:analysis*; D005445:Flunitrazepam / Q000032:analysis; D005506:Food Contamination / Q000032:analysis*; D006241:Haptens / Q000378:metabolism; D007118:Immunoassay; D007668:Kidney / Q000378:metabolism; D057230:Limit of Detection; D008099:Liver / Q000378:metabolism; D008460:Meat / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C; D009132:Muscles / Q000378:metabolism; D010076:Oxazepam / Q000032:analysis; D012709:Serum Albumin / Q000378:metabolism; D013552:Swine; D014018:Tissue Distribution
chemical_list: D000911:Antibodies, Monoclonal; D006241:Haptens; D012709:Serum Albumin; D001569:Benzodiazepines; D002998:Clonazepam; D005445:Flunitrazepam; D010076:Oxazepam; D003975:Diazepam
keywords: Benzodiazepine drugs; clonazepam; enzyme-linked immunosorbent assay; hapten; monoclonal antibody; swine tissues