Is Pichia pastoris a realistic platform for industrial production of recombinant human interferon gamma?

Razaghi, Ali, Tan, Emylin, Lua, Linda H.L., Owens, Leigh, Karhikeyan, O.P., and Heimann, Kirsten (2017) Is Pichia pastoris a realistic platform for industrial production of recombinant human interferon gamma? Biologicals, 45. pp. 52-60.

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Human interferon gamma (hIFNγ) is an important cytokine in the innate and adaptive immune system, produced commercially in Escherichia coli. Efficient expression of hIFNγ has been reported once for Pichia pastoris (Wang et al., 2014) – a proven heterologous expression system. This study investigated hIFNγ expression in P. pastoris replicating the previous study and expanding by using four different strains (X33: wild type; GS115: HIS−Mut+; KM71H: Arg+, Mut− and CBS7435: MutS) and three different vectors (pPICZαA, pPIC9 and pPpT4αS). In addition, the native sequence (NS) and two codon-optimised sequences (COS1 and COS2) for P. pastoris were used. Methanol induction yielded no expression/secretion of hIFNγ in X33, highest levels were recorded for CBS7435: MutS (∼16 μg. L−1). mRNA copy number calculations acquired from RT-qPCR for GS115-pPIC9-COS1 proved low abundance of mRNA. A 10-fold increase in expression of hIFNγ was achieved by lowering the minimal free energy of the mRNA and 100-fold by MutS phenotypes, substantially lower than reported by Wang et al. (2014). We conclude that commercial production of low cost, eukaryotic recombinant hIFNγ is not an economically viable in P. pastoris. Further research is required to unravel the cause of low expression in P. pastoris to achieve economic viability.

Item ID: 47271
Item Type: Article (Research - C1)
ISSN: 1095-8320
Keywords: biopharmaceuticals; Pichia pastoris; protein expression; interferon gamma; low-abundance RNA; minimum free energy
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A version of this publication was included as Chapter 3 of the following PhD thesis: Razaghi, Ali (2017) Evaluation of expression systems of recombinant human interferon gamma. PhD thesis, James Cook University, which is available Open Access in ResearchOnline@JCU. Please see the Related URLs for access.

Funders: Advanced Manufacturing Cooperative Research Centre (AMCRC), MBD Energy Research and Development programme for Biological Remediation of Methane from Underground Coal Mine Ventilation Air
Projects and Grants: AMCRC grant number 2.3.4, AMCRC PhD scholarship
Date Deposited: 19 Jun 2017 02:23
FoR Codes: 31 BIOLOGICAL SCIENCES > 3105 Genetics > 310505 Gene expression (incl. microarray and other genome-wide approaches) @ 50%
32 BIOMEDICAL AND CLINICAL SCIENCES > 3206 Medical biotechnology > 320603 Medical molecular engineering of nucleic acids and proteins @ 50%
SEO Codes: 86 MANUFACTURING > 8608 Human Pharmaceutical Products > 860803 Human Pharmaceutical Treatments (e.g. Antibiotics) @ 100%
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