A scalable low-cost cGMP process for clinical grade production of the HIV inhibitor 5P12-RANTES in Pichia pastoris.
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.
Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, Geneva, Switzerland.
Fabrice
F
Cerini
Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, Geneva, Switzerland.
Hubert
H
Gaertner
BioGrammatics Inc., Carlsbad, CA 92011, USA.
Knut
K
Madden
BioGrammatics Inc., Carlsbad, CA 92011, USA; Keck Graduate Institute, Claremont, CA, USA.
Ilya
I
Tolstorukov
Biologics Process Development, Inc., Poway, CA 92064, USA.
Scott
S
Brown
Department of Medical Protein Research, VIB-UGent, Ghent, Belgium; Department of Biochemistry and Microbiology, Laboratory for Protein Biochemistry and Biomolecular Engineering, Ghent University, Ghent, Belgium.
Bram
B
Laukens
Department of Medical Protein Research, VIB-UGent, Ghent, Belgium; Department of Biochemistry and Microbiology, Laboratory for Protein Biochemistry and Biomolecular Engineering, Ghent University, Ghent, Belgium.
Nico
N
Callewaert
University of Nebraska-Lincoln Biological Process Development Facility, College of Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Jay C
JC
Harner
University of Nebraska-Lincoln Biological Process Development Facility, College of Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Anna M
AM
Oommen
University of Nebraska-Lincoln Biological Process Development Facility, College of Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
John T
JT
Harms
University of Nebraska-Lincoln Biological Process Development Facility, College of Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Anthony R
AR
Sump
University of Nebraska-Lincoln Biological Process Development Facility, College of Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Robert C
RC
Sealock
University of Nebraska-Lincoln Biological Process Development Facility, College of Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Dustin J
DJ
Peterson
University of Nebraska-Lincoln Biological Process Development Facility, College of Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Scott K
SK
Johnson
LifeSci Partners, LLC, Vancouver, WA 98686-2811, USA.
Stephan B
SB
Abramson
University of Nebraska-Lincoln Biological Process Development Facility, College of Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Michael
M
Meagher
The Mintaka Foundation for Medical Research, Geneva, Switzerland.
Robin
R
Offord
Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, Geneva, Switzerland; The Mintaka Foundation for Medical Research, Geneva, Switzerland. Electronic address: oliver.hartley@unige.ch.
Oliver
O
Hartley
D019380:Anti-HIV Agents / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D019149:Bioreactors / Q000191:economics / Q000592:standards; D019742:Chemokines, CC / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005285:Fermentation; D006678:HIV / Q000187:drug effects*; D015658:HIV Infections / Q000188:drug therapy*; D006801:Humans; D020128:Inhibitory Concentration 50; D010843:Pichia; D010865:Pilot Projects; D053586:Virus Internalization / Q000187:drug effects
C539265:5P12-RANTES; D019380:Anti-HIV Agents; D019742:Chemokines, CC
5P12-RANTES; Biopharmaceutical; CCR5; Chemokine; HIV; Microbicide; Pichia; Process development; Yeast; cGMP