DF/HCC RESEARCH CORES

DF/HCC Recombinant Protein Expression and Purification Core
Cancer Pharmacology Core
Cytogenetics Core
High-Throughput Polymorphism Detection Core
Recombinant Protein Expression and Purification Core
Specialized Histopathology Core
Tissue MicroArray Core
Tumor Imaging Metrics Core
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Recombinant Protein Expression and Purification Core
Scientific Director: Dr. David Sachs
Location: Building CNY 149-6117C 13th St. Charlestown, MA 02129

 

Core Summary:

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/Contact Information:

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

Facilities & Equipment:

Location of Core:

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

Equipment:

Equipment
Manufacturer
BioFlo 510 Fermentor: 19.5L New Brunswick Scientific
HPLC System Shimadzu
Refrigerated Orbital Shaking Incubators (3) New Brunswick Scientific
Bio-Rad GenePulser X-Cell BioRad
Spectrophotometer BioRad
Large scale purification columns (Index 100/500 column) GE Healthcare
GeneAmp PCR System 9700 Applied Biosystem
Refrigerated Centrifuge 5417R Eppendorf
Refrigerated Centrifuge Legend RT Sorvall
The Endosafe-PTS Portable Test System Charles River Laboratories

 

Services:

Recombinant protein expression and purification in yeast Pichia pastoris

  • Codon optimization for yeast 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, SCF, GM-CSF) not available or prohibitively expensive from commercial sources

Cost effective small-scale production using E.coli or yeast Pichia pastoris expression system for a variety of applications including small animal studies, for use as immunogens, for ELISAs, and for high throughput microarray platforms

BioFlo 510 Fermentor (seen below) usage service

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

Fee Schedule/Rates:
Currently Available Products & Services
Internal Users
External Users
Partners & DF/HCC
Non-Partners AMC & Non-Profit
For-Profit & Industry
Murine IL2
$10/ 10ug
$14.4/ 10ug
Inquire
Human IL2
$10/ 10ug
$14.4/ 10ug
NHP CD3 immunotoxin
$900/ mg
$1,296/ mg
Porcine CD3 immunotoxin
$450/ mg
$648/ mg
Human Annexin A2
$90/ mg
$129.60/ mg
NHP IL3 (Gly and Non-N-Gly)
$90/ mg
$129.60/ mg
Porcine IL3 (Gly and Non-N-Gly)
Inquire
Cost + 44%
Porcine CTLA-4 DT390 fusion toxin
Porcine CTLA-4 (Gly and Non-N-Gly)
Murine CD40L (monomer, dimer, trimer)
Use of BioFlo 510 fermentor
New Recombinant Protein Development
$3,500 (see description)
$5,040
 
Recombinant Protein Development Service Description
Amount
Development of recombinant yeast expression protocol and purification strategy including:
$2,500
Codon optimization for expression in pichia pastoris system
Synthesis of entire codon optimized gene
Sequence confirmation
Cloning the gene into a yeast expression vector
Transformation of yeast and selection of transformants
Development of the expression protocol using shaker flasks
Verification of the expression by SDS gel and Western blot analysis
NOTE: If the expression level is too low or the protein cannot be solubilized, project may be terminated at this stage and investigator will be charged $2500 for service performed

Development of the two step purification protocol (can be used in vivo)

For convenient purification, we will add a 6xHis Tag in the C-terminus

$1,000
Purified material from 1L fermentation run using shaker flasks for functional testing
(No Charge)
Total
$3,500
The amount of material supplied depends on expression efficiency and yield after purification
Cost of additional protein to be determined based on expression level and yield

Getting Started:

To access the core, please contact Dr. Zhirui Wang at (617) 643-1957 or by email at
zhirui.wang@tbrc.mgh.harvard.edu.