Development and identification of monoclonal antibodies against ractopamine.
Ractopamine was reacted with two carrier proteins, human serum albumin and bovine thyroglobulin, as immunogen and coating antigen, respectively. Using a conventional immunization protocol, we generated a stable murine monoclonal antibody toward ractopamine, which had high affinities. The clone was found to be of IgG(2a) subclass with kappa light chain. An indirect competitive enzyme-linked immunosorbent assay for the determination of ractopamine has been optimized and characterized. The sensitivity, estimated as the IC(50) value, was 21.25 ng/mL, with a practical working range between 2.9 and 450 ng/mL. The limit of detection was 1.5 ng/mL. Moreover, other phenethanolamine beta-agonists showed low cross-reactivity with the monoclonal antibody. In addition, the indirect competitive enzyme-linked immunosorbent assay for the detection of ractopamine in animal feed was also developed using this antibody.
Li
Min
M
Department of Applied Chemistry, North University of China, Fen Xiao, Taiyuan, China. limin1701@sina.com
Chen
Zhimin
Z
Zhu
Qiufang
Q
D000318:Adrenergic beta-Agonists / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D000821:Animal Feed / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000915:Antibody Affinity; D002417:Cattle; D003429:Cross Reactions; D005260:Female; D005506:Food Contamination / Q000032:analysis; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000096:biosynthesis; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D010627:Phenethylamines / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D012709:Serum Albumin / Q000276:immunology; D013954:Thyroglobulin / Q000276:immunology
D000318:Adrenergic beta-Agonists; D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D010627:Phenethylamines; D012709:Serum Albumin; C060897:ractopamine; D013954:Thyroglobulin