The Recombinant Protein Expression and Purification (RPrEP) Core was established in 2009 to provide a cost-effective resource to the MGH and DF/ HCC community for expression and purification of recombinant proteins necessary for pre-clinical studies as well as for basic mechanistic studies by immunologists and cancer biologists.

Recombinant Proteins can be manufactured by this Core for a variety of applications according to MGH-DF/ HCC investigator needs. In addition to large-scale production of recombinant proteins in yeast Pichia pastoris, a cost effective small-scale production service will be provided using E.coli or yeast Pichia pastoris expression system (e.g. for small animal studies, use as immunogens, ELISAs, or high throughput microarray platforms).

Personnel:

• Scientific Director: Dr. David H. Sachs
• Scientific Co-director: Dr. Christene A. Huang
• Technical Director: Dr. Zhirui Wang
• Accounts Manager: Pat Kiszkiss

Contact Information:

Technical Director: Dr. Zhirui Wang
Phone: (617) 643-1957
Email: zhirui.wang@tbrc.mgh.harvard.edu

Location of Core:

The facility is located at CNY 149-6117C 13th St. Charlestown, MA 02129.

Equipment:

Recombinant protein expression and purification in yeast Pichia pastoris

• Codon optimization for yeast Pichia pastoris expression
• Synthesis of entire codon-optimized gene of interest
• Cloning/subcloning of the codon-optimized gene of interest into a yeast expression vector
• Transformation of yeast and selection of transformants
• Verification of expression of the gene of interest by SDS gel and Western blot
• Small-scale expression from one to three liters
• Development of efficient purification strategy
• Scale-up expression using BioFlo 510 fermentor

Large-scale purification of recombinant protein

Large-scale production of immunotoxins for cell depletion studies

Large-scale production of recombinant cytokines (e.g. IL-2, IL-3) not available or prohibitively expensive from commercial sources

Yeast display for protein-protein/antibody interaction study

BioFlo 510 Fermentor (seen below) training and usage

*Access to the Core will be available to all members of the research community. Priority will be given to MGH and DF/HCC users.  

Hermanrud CE, Lucas CL, Sykes M, Huang CA, Wang Z. (2011) Expression and purification of soluble murine CD40L monomers and polymers in yeast Pichia pastoris, Protein Expr Purif. 76:115-20.

Wang Z, Duran-Struuck R, Crepeau R, Matar A, Hanekamp I, Srinivasan S, Neville DM Jr, Sachs DH, Huang CA.(2011) Development of a diphtheria toxin based antiporcine CD3 recombinant immunotoxin, Bioconjug Chem. 22:2014-20.

Hermanrud CE, Pathiraja V, Matar A, Duran-Struuck R, Crepeau RL, Srinivasan S, Sachs DH, Huang CA, Wang Z. (2011) Expression and purification of non-N-glycosylated porcine interleukin 3 in yeast Pichia pastoris, Protein Expr Purif. In press.

Peraino J, Zhang H,  Hermanrud CE, Li G, Sachs DH, Huang CA, Wang Z. (2012) Expression and purification of soluble porcine CTLA-4 in yeast Pichia pastoris, Protein Expr Purif. In press.