Elsevier

MethodsX

Volume 7, 2020, 100769
MethodsX

Method Article
Protocol for bevacizumab purification using Ac-PHQGQHIGVSK-agarose

https://doi.org/10.1016/j.mex.2019.12.010Get rights and content
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Abstract

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.

Method name

Bevacizumab purification using Ac-PHQGQHIGVSK-agarose

Keywords

Peptide
Ligand
Downstream processing
Biopharmaceuticals
Monoclonal antibodies
Affinity chromatography
Solid phase peptide synthesis

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