Abstract

MIRANDA ARIZA, Alejandro; JUSTIZ LIMONTA, Anathais Carola; MURES DIAZ, Yany Leidys  and  ROMAGUERA FUENTES, Yisnelis. Obtention of cell culture supernatant for erythrocyte agglutination of blood group B antigen. Rev Cubana Hematol Inmunol Hemoter [online]. 2014, vol.30, n.3, pp. 249-256. ISSN 0864-0289.

Introduction : the obtention of monoclonal antibodies in ascite fluid has been declining gradually due to the appearance of alternative in vitro production that achieve higher volumes and a more precise monitoring of the production process, which increases the reproducibility of processes and the quality of products. Objective : to evaluate two methods to make cell culture supernatant rich in murine monoclonal IgG2b type, with agglutinating activity against human red cell of blood group antigen B (ABO system), which is secreted by murine hybridoma C6G4. Methods : two methods were evaluated for antibody production in cell culture supernatant using as model a CELLine bioreactor for the production of monoclonal antibodies in high cell density culture. Both methods essentially differed in the seeding cell density in the bioreactor and the fermentation period between seeding and harvesting of the culture broth rich in antibodies. The specific antibody concentration and potency of agglutination was determined in the obtained supernatant and also the cell density and cell viability of the culture reached at the time of harvest. Results : both methods generated culture supernatants with similar agglutination strength despite differences found in the rest of the variables measured. Even when one of the methods produced a higher antibody concentration in the supernatants, no differences in potency of the supernatants agglutination obtained by both alternatives were observed. Conclusions : both methods generated supernatant anti-B with different concentrations of antibodies but similar potency of agglutination. The main difference was that with one of the methods the same volume of the product was obtained in a considerably minor time.

Keywords : hybridoma; monoclonal antibody; in vitro production; CELLine bioreactor; hemagglutination.

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