Expression and purification of cGMP grade NY‐ESO‐1 for clinical trials

Lowe, Adam; Bardliving, Cameron; Huang, Chung‐Jr; Teixeira, Leonardo Maestri; Damasceno, Leonardo M.; Anderson, Kyle A.; Ritter, Gerd; Old, Lloyd J.; Batt, Carl A.

doi:10.1002/btpr.552

Cited by 11 publications

(7 citation statements)

References 57 publications

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“…Kim and coworkers have used pH-stat control to produce human glucagon-like peptide 1, resulting in a yield of 11.3 g/l [50]. It has also been used for the production of cancer vaccines, such as NY-ESO-1, Malan-A and SSX2, and showed high productivities [33,64]. This strategy, however, reXects the starvation of the cells instead of cell growth.…”

Section: Fed-batch Fermentation and Feeding Strategiesmentioning

confidence: 95%

Industrial production of recombinant therapeutics in Escherichia coli and its recent advancements

Huang

Lin

Yang

2012

Journal of Industrial Microbiology and Biotechnology

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372

248

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Nearly 30% of currently approved recombinant therapeutic proteins are produced in Escherichia coli. Due to its well-characterized genetics, rapid growth and high-yield production, E. coli has been a preferred choice and a workhorse for expression of non-glycosylated proteins in the biotech industry. There is a wealth of knowledge and comprehensive tools for E. coli systems, such as expression vectors, production strains, protein folding and fermentation technologies, that are well tailored for industrial applications. Advancement of the systems continues to meet the current industry needs, which are best illustrated by the recent drug approval of E. coli produced antibody fragments and Fc-fusion proteins by the FDA. Even more, recent progress in expression of complex proteins such as full-length aglycosylated antibodies, novel strain engineering, bacterial N-glycosylation and cell-free systems further suggests that complex proteins and humanized glycoproteins may be produced in E. coli in large quantities. This review summarizes the current technology used for commercial production of recombinant therapeutics in E. coli and recent advances that can potentially expand the use of this system toward more sophisticated protein therapeutics.

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Section: Fed-batch Fermentation and Feeding Strategiesmentioning

confidence: 95%

Industrial production of recombinant therapeutics in Escherichia coli and its recent advancements

Huang

Lin

Yang

2012

Journal of Industrial Microbiology and Biotechnology

Self Cite

372

248

View full text Add to dashboard Cite

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“…NY-ESO-1 protein for immunization was manufactured at the Ludwig Institute for Cancer Research (LICR) as described (26). Incomplete Freund’s adjuvant, IFA (Montanide ™ ISA-51), contained mineral oil (Drakeol) and anhydro mannitol octadecanoate and was manufactured by Seppic Inc. Sargramostim, (Leukine®) was obtained from Bayer (Seattle, WA).…”

Section: Methodsmentioning

confidence: 99%

Epigenetic Potentiation of NY-ESO-1 Vaccine Therapy in Human Ovarian Cancer

Odunsi

Matsuzaki

James

et al. 2014

Cancer Immunology Research

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The cancer-testis/cancer-germline antigen NY-ESO-1 is a vaccine target in epithelial ovarian cancer (EOC), but its limited expression is a barrier to vaccine efficacy. As NY-ESO-1 is regulated by DNA methylation, we hypothesized that DNA methyltransferase (DNMT) inhibitors may augment NY-ESO-1 vaccine therapy. In agreement, global DNA hypomethylation in EOC was associated with the presence of circulating antibodies to NY-ESO-1. Pre-clinical studies using EOC cell lines showed that decitabine treatment enhanced both NY-ESO-1 expression and NY-ESO-1-specific CTL-mediated responses. Based on these observations, we performed a phase I dose-escalation trial of decitabine, as an addition to NY-ESO-1 vaccine and doxorubicin liposome (doxorubicin) chemotherapy, in 12 patients with relapsed EOC. The regimen was safe, with limited and clinically manageable toxicities. Both global and promoter-specific DNA hypomethylation occurred in blood and circulating DNAs, the latter of which may reflect tumor cell responses. Increased NY-ESO-1 serum antibodies and T cell responses were observed in the majority of patients, and antibody spreading to additional tumor antigens was also observed. Finally, disease stabilization or partial clinical response occurred in 6/10 evaluable patients. Based on these encouraging results, evaluation of similar combinatorial chemo-immunotherapy regimens in EOC and other tumor types is warranted.

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“…The average diameter of the NoV VLPs in the bioreactor (>60 nm) could be explained by the high salt concentration in the media, which has been shown to affect the particle size, as described with HPV VLPs expression in P. pastoris [41]. Aggregation of the VLPs could be prevented through lyophilization, the use of detergents, sorbitol, or other buffers [20,41,42]. …”

Section: Discussionmentioning

confidence: 99%

Secreted production of assembled Norovirus virus-like particles from Pichia pastoris

et al. 2014

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BackgroundNorovirus virus-like particles (NoV VLPs) have recently been explored as potential vaccine platforms due to their ability to produce an effective immune response. Expression of the main structural protein, VP1, leads to formation of self-assembled particles with similar characteristics to the original virus. These NoV VLPs have been expressed in Escherichia coli, yeast and insect cells. Expression in E. coli and insect cells share downstream processing issues due to the presence of inclusion bodies or the need for numerous purification steps. NoV VLPs have also been produced in the yeast P. pastoris; however the protein was only expressed intracellularly.ResultsWe have successfully expressed and secreted the VP1 protein in the novel P. pastoris strain, Bg11, using the methanol inducible pJ912 expression vector, containing the cDNA of NoV VP1. Expression of the VP1 protein in Bg11 was carried out in a 1.5 L bioreactor resulting in a total yield of NoV VLPs greater than 0.6 g/L. NoV VLPs obtained from the culture supernatant were purified via ion-exchange chromatography, resulting in particles with a purity over 90%. The average size of the particles after purification was 40 nm. Transmission electron microscopy was used to visualize the morphology of the particles and saliva-binding assay confirmed that the NoV VLPs bind to Histo-Blood Group Antigens (HBGA).ConclusionsIn this study we describe the expression and characterization of fully assembled Norovirus virus-like particles obtained from P. pastoris. The particles are similar in size, morphology and binding capacity, as previously described, for the original NoV. Our results detail the successful expression and secretion of VLPs in P. pastoris, improving their candidacy as a vaccine platform.

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Expression and purification of cGMP grade NY‐ESO‐1 for clinical trials

Cited by 11 publications

References 57 publications

Industrial production of recombinant therapeutics in Escherichia coli and its recent advancements

Industrial production of recombinant therapeutics in Escherichia coli and its recent advancements

Epigenetic Potentiation of NY-ESO-1 Vaccine Therapy in Human Ovarian Cancer

Secreted production of assembled Norovirus virus-like particles from Pichia pastoris

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