A scalable low-cost cGMP process for clinical grade production of the HIV inhibitor 5P12-RANTES in Pichia pastoris

Authors

Fabrice Cerini, University of Geneva, Geneva, Switzerland
Hubert Gaertner, University of Geneva, Geneva, Switzerland
Knut Madden, BioGrammatics Inc., Carlsbad, CA
Ilya Tolstorukov, BioGrammatics Inc., Carlsbad, CA
Scott Brown, Biologics Process Development, Inc
Bram Laukens, Department of Medical Protein Research, VIB-UGent, Ghent, Belgium
Nico Callewaert, Unit for Molecular GlycobiologyFollow
Jay C. Harner, University of Nebraska-LincolnFollow
Anna M. Oommen, University of Nebraska - LincolnFollow
John T. Harms, University of Nebraska - LincolnFollow
Anthony R. Sump, University of Nebraska - LincolnFollow
Robert C. Sealock, University of Nebraska - LincolnFollow
Dustin J. Peterson, University of Nebraska - LincolnFollow
Scott K. Johnson, University of Nebraska - LincolnFollow
Stephan B. Abramson, LifeSci Partners, LLC, Vancouver
Michael M. Meagher, University of Nebraska - LincolnFollow
Robin Offord, The Mintaka Foundation for Medical Research, Geneva, Switzerland
Oliver Hartley, University of Geneva, Geneva, SwitzerlandFollow

Date of this Version

2016

Document Type

Article

Citation

Protein Expression and Purification 119 (2016) 1-10.
doi 10.1016/j.pep.2015.10.01

Comments

Copyright 2015 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license.

Abstract

In the continued absence of an effective anti-HIV vaccine, approximately 2 million new HIV infections occur every year, with over 95% of these in developing countries. Calls have been made for the development of anti-HIV drugs that can be formulated for topical use to prevent HIV transmission during sexual intercourse. Because these drugs are principally destined for use in low-resource regions, achieving production costs that are as low as possible is an absolute requirement. 5P12-RANTES, an analog of the human chemokine protein RANTES/CCL5, is a highly potent HIV entry inhibitor which acts by achieving potent blockade of the principal HIV coreceptor, CCR5. Here we describe the development and optimization of a scalable low-cost production process for 5P12-RANTES based on expression in Pichia pastoris. At pilot (150 L) scale, this cGMP compliant process yielded 30 g of clinical grade 5P12- RANTES. As well as providing sufficient material for the first stage of clinical development, this process represents an important step towards achieving production of 5P12-RANTES at a cost and scale appropriate to meet needs for topical HIV prevention worldwide.