Integrated Separation And Purification Ofβ-hCG Monoclonal Antibody

β-hCG monoclonal antibody has been widely used in the pregnancy and pregnancy-related disease diagnosis. Therefore, the preparation of higher-purity β-hCG antibody has became the driving force in the current research. Affinity aqueous two-phase extraction as a new promising separation approach has been used in the purification of bioactivators, which combines aqueous two-phase extraction with affinity separation.In this paper, the partitioning of β-hCG monoclonal antibody in polyethylene glycol(PEG)/trisodium citrate affinity aqueous two-phase systems(ATPS) using sulfamethazine(SM2) as affinity ligand was investigated. Furthermore, effects of PEG molecular, PEG component (PEG4000, PEG4000-SM2), trisodium citrate concentration, pH and (3-hCG antibody concentration were studied as well. The optimal conditions were PEG40009%, PEG4000-SM28%, trisodium citrate12%, pH6.0and β-hCG antibody0.5mg/g. Under above conditions, the highest Ke of7.6and Ye of91.1%were obtained, the activity of β-hCG monoclonal antibody stayed the same as before.Based on the above results, extraction of β-hCG monoclonal antibody direct form mouse ascites using affinity ATPS was investigated. The results showed that purity of β-hCG monoclonal antibody was relatively low (P=36.8%) with single extraction and the extraction stage has no improvement on it. However, while employed5%PEG10000/8%trisodium citrate ATPS to pretreat the mouse ascites and then rebuilt the previously optimized affinity ATPS to extract β-hCG monoclonal antibody, the purity of82.1%, the yield of81.7%were obtained and the purification factor was5.3.