title: [Separation strategy of affinity chromatography for mixed antibodies].
abstract: A mammary gland bioreactor can efficiently express human recombinant monoclonal antibody. However, the target products are similar to the bovine antibody in the raw emulsion material in properties and structures. Thus it is difficult to achieve effective separation of the target products. In this work, the species differences between bovine antibody and recombinant human antibody were analyzed and a new separation strategy was raised based on it. We employed two kinds of affinity chromatography to separate these two antibodies from each other and studied the effect of elution mode upon separation. The results demonstrated that Protein A affinity chromatography could get hybrid antibodies using gradient elution mode, but hardly separate the recombinant human antibody and bovine antibody from each other. In contrast, the combination of Protein A affinity chromatography and displacement chromatography could separate the hybrid antibodies effectively and finally give recombinant human IgG (rHGG) product with the purity of 95% and the yield of more than 95%. Immuno-affinity chromatography could also effectively purify recombinant monoclonal antibodies and owned better generality, which could be used in purification of recombinant antibody expressed by any animal mammary gland. -
authors.affiliation: Institute of Life Science and Technology, Kunming University of Science and Technology, Kunming 650224, China.
authors.forename: Yuefang
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Luo
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Limei
authors.initials: L
authors.lastname: Kang
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Shi
authors.affiliation: Institute of Life Science and Technology, Kunming University of Science and Technology, Kunming 650224, China.
authors.forename: Qingxiong
authors.initials: Q
authors.lastname: Meng
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Su
mesh_terms: D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D002417:Cattle; D002846:Chromatography, Affinity; D006801:Humans; D007074:Immunoglobulin G; D011994:Recombinant Proteins
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011994:Recombinant Proteins