Characterization and Scale-Up of Cyanogen Bromide Chemical Activation of Sepharose CL-4B in a Stirred Tank Reactor to Purify the rHBsAg

Eutimio G. Fernández, Center for Engineering and Chemical Researches
José A. Montero, Center for Genetic Engineering and Biotechnology
Rodolfo Valdés, Center for Genetic Engineering and Biotechnology
Milagros G. Arce, Center for Engineering and Chemical Researches
Andrés B. Tamayo, Center for Genetic Engineering and Biotechnology
Déborah Geada, Tobacco Research Institute
Yenisley Medina, Center for Genetic Engineering and Biotechnology
Malvina Benitez, Center for Engineering and Chemical Researches
Yarysel Guevara, Center for Genetic Engineering and Biotechnology
William Ferro, Center for Genetic Engineering and Biotechnology
Jorge L. López, Center for Genetic Engineering and Biotechnology
Maylin La O, Center for Genetic Engineering and Biotechnology
Yuirisleyi Aldama, Center for Genetic Engineering and Biotechnology
Sigifredo A. Padilla, Center for Genetic Engineering and Biotechnology
Alejandro Figueroa, Center for Genetic Engineering and Biotechnology

Abstract

The aim of this paper was to characterize and to scale-up the CNBr chemical activation of Sepharose CL-4B in a stirred tank reactor to purify the recombinant Hepatitis B surface antigen for use in vaccines. The activation quality was assessed measuring the concentration of the cyanate esters and the percentage of nitrogen in the activated matrix. The Sepharose CL-4B incorporated less than 50% of the initial CNBr amount added to the matrix, but the monoclonal antibody CB.Hep-1, used as immunoligand to purify the rHBsAg, was efficiently immobilized in a weakly activated matrix (<8 μmol/mL of matrix). The impellent type has a significant influence on activated matrix quality. The constant power per volume unit was a proper criterion to scale-up the Sepharose CL-4B March's modified chemical activation procedure with CNBr.

Recommended Citation

Fernández, Eutimio G.; Montero, José A.; Valdés, Rodolfo; Arce, Milagros G.; Tamayo, Andrés B.; Geada, Déborah; Medina, Yenisley; Benitez, Malvina; Guevara, Yarysel; Ferro, William; López, Jorge L.; La O, Maylin; Aldama, Yuirisleyi; Padilla, Sigifredo A.; and Figueroa, Alejandro (2008) "Characterization and Scale-Up of Cyanogen Bromide Chemical Activation of Sepharose CL-4B in a Stirred Tank Reactor to Purify the rHBsAg," International Journal of Chemical Reactor Engineering: Vol. 6: A33.
Available at: http://www.bepress.com/ijcre/vol6/A33

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