Purification of a Divalent Version of Antibody Fragments Specific for a Novel Epitope of the Human Vascular Endothelial Growth Factor with Low Aggregate Level and High Purity

by Hasel Aragón, Lincidio Pérez, Dayron Ojeda, Daily Hernández, Sigifredo Padilla, Marcos González, Jorge Gavilondo, Marta Ayala, Alexis Mussachio, Mónica Bequet, Humberto Lamdan, Regla Somoza, Mayda Candelario, Andrés Tamayo, Cristina García, Gisela Calas, Yanara González, Adelma Pérez, and Rodolfo Valdés
Volume 15, Issue 4 (Winter 2016/2017)

Cancer is one of the leading causes of death worldwide, and the second leading cause of death in Cuba. To address this serious health problem, some research has involved suppressing tumor growth by inhibiting the angiogenesis process using several molecules including antibodies. A divalent version of antibody fragments, the CIGB-598a, with a molecular weight between 100 and 110 kDa, has been expressed in CHO cells specific for a novel epitope of the human vascular endothelium growth factor (VEGF). This material has been generated at the Center for Genetic Engineering and Biotechnology to support cancer research efforts. As in other studies involving the purification of recombinant molecules, CIGB-598a exhibited a high degree of aggregation in the CHO cell culture supernatant. This required the design of a downstream process capable of removing high levels of aggregates to obtain a highly pure target molecule for use in preclinical studies and human applications further down the road. We have developed a suitable downstream method based on the combination of three chromatography processes: affinity, cation-exchange, and anion-exchange that recover a relatively low level of CIGB-598a, but at a level of high purity (greater than 95 %) with fewer aggregates (below 1%)…

Arag√≥n H, P√©rez L, Ojeda D, Hern√°ndez D, Padilla S, Gonz√°lez M et al. Purification of a divalent version of antibody fragments specific for a novel epitope of the human vascular endothelial growth factor with low aggregate level and high purity. BioProcess J, 2017; 15(4): 32‚Äď41.¬†https://doi.org/10.12665/J154.Valdes.P

Posted online February 15, 2017.