Hydrophobic charge-induction resin with 5-aminobenzimidazol as the functional ligand: preparation, protein adsorption and immunoglobulin G purification.
A new hydrophobic charge-induction chromatography resin was prepared with 5-aminobenzimidazol as functional ligand and polyacrylic ester beads as matrix. Adsorption isotherms and adsorption in columns were investigated using human immunoglobulin G and bovine serum albumin as model proteins, and the influence of pH and NaCl concentration was discussed. Results showed that the ligand density was 195 μmol/mL gel, and protein selectivity can be improved by controlling pH and salt addition. An optimized purification process (sample loading at pH 8.0 with 0.2 M NaCl and elution at pH 5.0) was performed to purify human immunoglobulin G from bovine serum albumin containing feedstock, which resulted in human immunoglobulin G purity of 99.7% and recovery of 94.6%. A similar process was applied for the purification of monoclonal antibody from cell culture supernatant, which showed antibody purity of 94.9% and recovery of 92.5%. The results indicated that the new resin developed had comparable performance as Protein A chromatography and would be suitable for antibody purification from complex feedstock.
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
Jun
J
Yan
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
Qi-Lei
QL
Zhang
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
Hong-Fei
HF
Tong
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
Dong-Qiang
DQ
Lin
Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
Shan-Jing
SJ
Yao
D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001562:Benzimidazoles / Q000032:analysis* / Q000737:chemistry*; D016466:CHO Cells; D002417:Cattle; D002845:Chromatography / Q000295:instrumentation*; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000737:chemistry* / Q000302:isolation & purification; D008024:Ligands; D012492:Salts; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry
D000911:Antibodies, Monoclonal; D001562:Benzimidazoles; D007074:Immunoglobulin G; D008024:Ligands; D012492:Salts; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; C059092:5-aminobenzimidazole
Adsorption; Bovine serum albumin; Human immunoglobulin G; Hydrophobic charge-induction chromatography; Purification