Protocol for bevacizumab purification using Ac-PHQGQHIGVSK-agarose

BARREDO, GABRIELA R.; GIUDICESSI, SILVANA L.; MARTÍNEZ CERON, MARÍA C.; SAAVEDRA, SOLEDAD L.; RODRÍGUEZ, SANTIAGO; FILGUEIRA RISSO, LUCAS; ERRA-BALSELLS, ROSA; MAHLER, GUSTAVO; ALBERICIO, FERNANDO; CASCONE, OSVALDO; CAMPERI, SILVIA A.

MethodsX

Elsevier B.V.

Año: 2020 vol. 7

2215-0161

Bevacizumab is a monoclonal antibody, produced in CHO cells, used for the treatment of many human cancers. It is an anti-vascular endothelial growth factor (antsi-VEGF) that blocks the growth of tumor blood vessels. Nowadays its purification is achieved by affinity chromatography (AC) using protein A which is a very expensive ligand. On the other hand, the peptide Ac-PHQGQHIGVSK contained in the VEGF fragment binds bevacizumab with high affinity. This short peptide ligand has higher stability and lower cost than protein A and it can be prepared very easily by solid phase peptide synthesis. The present protocol describes the synthesis of AcPHQGQHIGVSK-agarose and its use for affinity chromatography purification of bevacizumab from a clarified CHO cell culture.