Stirrer tank: an appropriate technology to immobilize the CB.Hep-1 monoclonal antibody for immunoaffinity purification |
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Institution: | 1. Department of Chemistry and Polymer, Faculty of Chemistry and Petrochemical Engineering, Standard Research Institute, P.O. Box: 31745-139, Karaj, Iran;2. Department of Chemistry, Payame Noor University, P.O. BOX 19395-3697, Tehran, Iran;1. Institute of Business Scientific, Henan Academy of Sciences, Wenhua Road #87, Zhengzhou, Henan 450003, PR China;2. Zhengzhou Tobacco Research Institute of CNTC, Fengyang Street #2, Zhengzhou, Henan 450001, PR China |
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Abstract: | The CB.Hep-1 monoclonal antibody was coupled to CNBr-activated Sepharose CL 4B at three different immobilization scales for purification of recombinant hepatitis B surface antigen. Standard laboratory apparatus to obtain immunosorbents of 1 l (scale I) and 3 l (scale II) as well as a stirrer tank to prepare 6 l immunosorbents (scale III) were used. The binding capacity at scale III was 2- and 1.5-fold higher with respect to the scales II and I, while a reduction in the ligand leakage of 5- and 2-folds was observed. Immunosorbents from scale II showed a significantly reduced adsorption, and an increased ligand leakage. Differences in the coupling efficiency were not observed. Antigen purity eluted from the immunosorbents was always above 85%. |
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