title: [Purification of anti-HBcAg monoclonal antibodies using immobilized metal ion affinity chromatography].
abstract: Anti-HBcAg monoclonal antibodies from mouse ascites were purified by using immobilized metal ion affinity chromatography. We optimized the conditions of sample loading and elution. The results showed that when the pH stepwise elution was used, the best solution for sample loading was 20 mmol/L phosphate buffer containing 0.5 mol/L sodium chloride at pH 8.0 and the mAb was eluted at pH 5.0. The purity of obtained mAb was more than 85% and recovery reached 80%. When the adsorbed proteins were eluted by using gradient elution of an imidazole, the best solution for loading condition was 20 mmolL phosphate buffer containing 5 mmol/L imidazole at pH 8.0. The purity and recovery of antibody were up to 95%.
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, China.
authors.forename: Ji
authors.initials: J
authors.lastname: Zhu
authors.forename: Yu
authors.initials: Y
authors.lastname: Yi
authors.forename: Yinfei
authors.initials: Y
authors.lastname: Wu
authors.forename: Keyin
authors.initials: K
authors.lastname: Zhu
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Mei
authors.forename: Jianshu
authors.initials: J
authors.lastname: Chen
authors.forename: Guoqing
authors.initials: G
authors.lastname: Ying
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D006512:Hepatitis B Core Antigens / Q000276:immunology*; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000737:chemistry; D008670:Metals / Q000737:chemistry; D051379:Mice
chemical_list: D000911:Antibodies, Monoclonal; D006512:Hepatitis B Core Antigens; D007093:Imidazoles; D008670:Metals; C029899:imidazole