A short peptide fragment of the vascular endothelial growth factor as a novel ligand for bevacizumab purification.

Autor: Barredo GR; Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956, 1113, Buenos Aires, Argentina; CONICET-Universidad de Buenos Aires, Instituto de Nanobiotecnología (NANOBIOTEC), Facultad de Farmacia y Bioquímica, Junín 956, 1113, Buenos Aires, Argentina., Giudicessi SL; Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956, 1113, Buenos Aires, Argentina; CONICET-Universidad de Buenos Aires, Instituto de Nanobiotecnología (NANOBIOTEC), Facultad de Farmacia y Bioquímica, Junín 956, 1113, Buenos Aires, Argentina., Martínez Ceron MC; Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956, 1113, Buenos Aires, Argentina; CONICET-Universidad de Buenos Aires, Instituto de Nanobiotecnología (NANOBIOTEC), Facultad de Farmacia y Bioquímica, Junín 956, 1113, Buenos Aires, Argentina., Saavedra SL; Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956, 1113, Buenos Aires, Argentina; CONICET-Universidad de Buenos Aires, Instituto de Nanobiotecnología (NANOBIOTEC), Facultad de Farmacia y Bioquímica, Junín 956, 1113, Buenos Aires, Argentina., Rodriguez S; MAbxience SAU, Carlos Villate 5148, 1605, Munro, Buenos Aires, Argentina., Filgueira Risso L; MAbxience SAU, Carlos Villate 5148, 1605, Munro, Buenos Aires, Argentina., Erra-Balsells R; Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Química Orgánica, Pabellón II. Ciudad Universitaria, 1428, Buenos Aires, Argentina; CONICET, Universidad de Buenos Aires, Centro de Investigación en Hidratos de Carbono (CIHIDECAR), Facultad de Ciencias Exactas y Naturales, Pabellón II Ciudad Universitaria, 1428, Buenos Aires, Argentina., Mahler G; AGC Biologics, 22021 20th Avenue SE, Bothell, WA, 98021, USA., Albericio F; School of Chemistry & Physics, University of KwaZulu-Natal, Durban, 4001, South Africa; CIBER-BBN, Networking Centre on Bioengineering, Biomaterials and Nanomedicine, Department of Organic Chemistry, University of Barcelona, Barcelona, 08028, Spain., Cascone O; Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956, 1113, Buenos Aires, Argentina; CONICET-Universidad de Buenos Aires, Instituto de Nanobiotecnología (NANOBIOTEC), Facultad de Farmacia y Bioquímica, Junín 956, 1113, Buenos Aires, Argentina., Camperi SA; Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Junín 956, 1113, Buenos Aires, Argentina; CONICET-Universidad de Buenos Aires, Instituto de Nanobiotecnología (NANOBIOTEC), Facultad de Farmacia y Bioquímica, Junín 956, 1113, Buenos Aires, Argentina. Electronic address: scamperi@ffyb.uba.ar.
Jazyk: angličtina
Zdroj: Protein expression and purification [Protein Expr Purif] 2020 Jan; Vol. 165, pp. 105500. Date of Electronic Publication: 2019 Sep 19.
DOI: 10.1016/j.pep.2019.105500
Abstrakt: Bevacizumab is a vascular endothelial growth factor (VEGF)-directed monoclonal antibody (mAb) used for the treatment of several human cancers. Given that bevacizumab is administered intravenously, it must have extremely high purity, which is achieved by purification with protein A affinity chromatography (AC). However, protein A is a very expensive ligand, thereby increasing the cost of purification. Furthermore, the harsh elution conditions required to recover bevacizumab from the AC column can damage both the mAb and protein A. In contrast, short peptides show higher stability, easier synthesis and lower cost and are therefore ideal ligands for AC. In the present study, the peptide Ac-PHQGQHIGVSK contained in the VEGF fragment that binds bevacizumab, was synthesized and immobilized on agarose. The peptidyl-agarose showed affinity for bevacizumab, with an equilibrium dissociation constant value of 2.2±0.5 x 10 -7  M under optimal conditions. Samples of CHO cell filtrate producing bevacizumab were loaded on the peptidyl-agarose chromatography column. Bevacizumab was recovered from the elution fraction with a yield of 94% and a purity of 98%. The maximum capacity (qm) 38±2 mg of bevacizumab per mL of matrix was comparable to that of commercial protein A matrices. Moreover, the peptide ligand showed greater stability and a lower cost than protein A. Unlike peptides previously reported for IgG purification, the ligand described herein allows mAb elution under mild conditions, thereby favoring the integrity of bevacizumab. The lack of Trp, Met or Cys in the peptide prevents its oxidation and extends the useful life of the chromatographic matrix.
(Copyright © 2019 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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