Protocol for bevacizumab purification using Ac-PHQGQHIGVSK-agarose

MethodsX

Gustavo Mahler¹  Lucas Filgueira Risso¹  Soledad L. Saavedra¹  Silvana L. Giudicessi¹  Silvia A. Camperi²  Rosa Erra-Balsells³  María C. Martínez Ceron³  Gabriela R. Barredo³  Santiago Rodríguez³  Osvaldo Cascone⁴  Fernando Albericio⁴ 

[1]CONICET-Universidad de Buenos Aires, Instituto de Nanobiotecnología (NANOBIOTEC), Facultad de Farmacia y Bioquímica, Junín 956, 1113, Buenos Aires, Argentina

[2]Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Orgánica, Pabellón II, Ciudad Universitaria, 1428, Buenos Aires, Argentina

[3]Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956, 1113, Buenos Aires, Argentina

[4]mAbxience SAU, Carlos Villate 5148, 1605, Munro, Buenos Aires, Argentina

关键词: Peptide;    Ligand;    Downstream processing;    Biopharmaceuticals;    Monoclonal antibodies;    Affinity chromatography;   

DOI  :  

来源: DOAJ

【 摘 要 】

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 Ac-PHQGQHIGVSK-agarose and its use for affinity chromatography purification of bevacizumab from a clarified CHO cell culture. • Ac-PHQGQHIGVSK-agarose capacity and selectivity are equivalent to those of protein A matrices. • The peptide ligand shows a greater stability and lower cost. The lack of Trp, Met or Cys in the peptide ligand prevents its oxidation and extends the useful life of the chromatographic matrix. • Mild conditions used during chromatography preserved the integrity of bevacizumab.

【 授权许可】

Unknown