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Comparison of Sepharose CL-2B and CL-4B Matrices in Purification of the Hepatitis B Surface Antigen by Affinity Chromatography |
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| Authors: | Eutimio G Fernández Rodolfo Valdés Yenisley Medina José A Montero Alejandro Figueroa Williams Ferro Yarysel Guevara Tatiana Álvarez Andrés Tamayo Regla M Somoza Leonardo Canaán-Haden Jorge L López |
| Institution: | 1. Department of Inorganic Chemistry, Center for Engineering and Chemical Researches (CIIQ), F Street, #115 be/Ave 5th and Calzada, 10400, Havana, Cuba 2. Department of Monoclonal Antibody Production, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Ave 31 be/158 and 190, Playa, 10600, Havana, Cuba 3. Department of Process Control, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Ave 31 be/158 and 190, Playa, 10600, Havana, Cuba 4. Department of Chemistry-Physcis, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Ave 31 be/158 and 190, Playa, 10600, Havana, Cuba 5. Direction of Quality Control, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Ave 31 be/158 and 190, Playa, 10600, Havana, Cuba |
| Abstract: | Sepharose CL-4B and Sepharose CL-2B with immobilized CB.Hep-1 monoclonal antibody were compared for studying the matrix efficiency in the immunopurification of the recombinant hepatitis B surface antigen (rHBsAg). The elution capacities of both matrices were similar over eight chromatographic cycles, significant differences were only observed in the first purification cycles. A high percentage of the adsorbed rHBsAg was not eluted from both matrices. The rHBsAg purity was not affected by matrix characteristics (pore size and percentage of agarose) and differences between Sepharose CL-4B and Sepharose CL-2B do not provoke differences in the antibody released from the matrices under defined experimental conditions. |
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