title: Antibody purification using affinity chromatography: a case study with a monoclonal antibody to ractopamine.
abstract: The application of antibodies to small molecules in the field of bioanalytics requires antibodies with stable biological activity and high purity; thus, there is a growing interest in developing rapid, inexpensive and effective procedures to obtain such antibodies. In this work, a ractopamine (RAC) derivative, N-4-aminobutyl ractopamine (ABR), was synthesized for preparing new specific affinity chromatography to purify a murine monoclonal antibody (mAb) against RAC from ascites. The performance of the new specific chromatography was compared with four other purification methods in terms of recovery, purity and biological activity of mAb. These four purification methods were prepared by using specific ligands (RAC and RAC-ovalbumin) and commercial ligands (protein G and protein A), respectively. The results showed that the highest recovery (88.1%) was achieved using the new chromatography; in comparison, the recoveries from the other methods were all below 70%. The purity of the mAbs from the new chromatography was 88.3%, while, the highest purity of 97.6% was from protein G chromatography and the lowest purity of 84.7% was from protein A chromatography. The biological activity of the purified mAb from all of the chromatography methods was comparable in enzyme-linked immunosorbent immunoassay (ELISA).
authors.affiliation: College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China; Beijing Laboratory for Food Quality and Safety, Beijing 100193, People's Republic of China.
authors.forename: Zhanhui
authors.initials: Z
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Liang
authors.affiliation: College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China; Beijing Laboratory for Food Quality and Safety, Beijing 100193, People's Republic of China.
authors.forename: Kai
authors.initials: K
authors.lastname: Wen
authors.affiliation: College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China; Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing 100193, People's Republic of China.
authors.forename: Suxia
authors.initials: S
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China; National Reference Laboratories for Veterinary Drug Residue, Beijing 100093, People's Republic of China. Electronic address: sjz@cau.edu.cn.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Shen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D000918:Antibody Specificity; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D010047:Ovalbumin; D010627:Phenethylamines / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D010627:Phenethylamines; C060897:ractopamine; D010047:Ovalbumin
keywords: Antibody; Chromatography; Ligand; Purification; Ractopamine; Small molecule