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

Adam J Lowe1, Cameron L Bardliving, Chung-Jr Huang

  • 1Graduate Field of Microbiology, Cornell University, Ithaca, NY 14853, USA. ajl248@cornell.edu

Biotechnology Progress
|March 3, 2011
PubMed

Insights

Clinical-grade NY-ESO-1 cancer vaccine production was achieved using Good Manufacturing Practices. This His-tagged protein, expressed in E. coli, yielded over 3.4 g with minimal endotoxin for Phase I trials.

Area of Science:

  • Oncology
  • Biotechnology
  • Immunology

Background:

  • NY-ESO-1 is a cancer-testis antigen found in various cancers.
  • NY-ESO-1 shows promise as a cancer vaccine, activating the immune system against tumors.

Purpose of the Study:

  • Develop and execute a Current Good Manufacturing Practices (cGMP) compliant process.
  • Produce clinical-grade NY-ESO-1 protein for Phase I clinical trials.

Main Methods:

  • Expressed His-tagged NY-ESO-1 in C41DE3 Escherichia coli using a T-7 promoter.
  • Utilized a 20 L fed-batch fermentation with pH-stat control.
  • Purified the protein from inclusion bodies via a three-column chromatography process.

Main Results:

  • Achieved a high yield of over 3.4 grams of purified NY-ESO-1 protein.
  • Ensured endotoxin levels were below 0.005 EU/μg, meeting stringent clinical standards.
  • Successfully produced clinical-grade material suitable for human trials.

Conclusions:

  • A robust cGMP process was established for large-scale NY-ESO-1 production.
  • The developed method ensures high yield and purity of the cancer vaccine candidate.
  • This facilitates the advancement of NY-ESO-1 into clinical evaluation for cancer treatment.

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