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New Core: Recombinant Protein Expression and Purification

Fig. 1: Large Scale BioFlo 510 Fermentor

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

Recombinant proteins may be manufactured for a variety of applications according to 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 yeast pichia pastoris and/or bacteria expression system (e.g. for small animal studies, use as immunogens, ELISAs, or high-throughput microarray platforms).  

The new BioFlo 510 Fermentor (Fig. 1) allows the core to produce large scale recombinant proteins using yeast pichia pastoris system.

Currently available recombinant protein products in the core:  

  • Anti-monkey CD3 immunotoxin
  • Anti-porcine CD3 immunotoxin
  • Porcine IL3
  • Monkey IL3
  • DT390-Porcine CTLA4
  • Mouse CD40L (monomer, dimer, trimer)
  • Human annexin A2
  • Human IL2   

 

Products are priced to recover costs of production only and vary depending on the expression efficiency and yield for each particular recombinant protein following purification.  Purified products are quality controlled for in vivo use. Recombinant anti-monkey CD3 immunotoxin, available for use to deplete the monkey CD3 T cells in vivo, is shown below (Fig. 2) as an example of purified product available from the core.


Fig. 2. Anti-monkey CD3 immunotoxin: A-dmDT390-scfbDb(C207), (97.7 kDa). Lane 1: See Blue Plus2 Prestained Standard (Invitrogen); Lane 2: 0.6 μg of anti-monkey CD3 immunotoxin, Lot# ZW14; Lane 3: 1.2 μg of anti-monkey CD3 immunotoxin, Lot# ZW14.

To access the core's services please contact Dr. Zhirui Wang @ 617-643-1957 or email: zhirui.wang@tbrc.mgh.harvard.edu.