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Protein Expression and Production

Protein Expression Laboratory

The Protein Expression Laboratory develops, improves, and delivers protein-centric services. Our goal is to help client investigators achieve their research goals with the lowest possible cost in the shortest time. All PEL services are available individually as well as part of larger projects.

The PEL Protein Pipeline

The PEL has constructed hundreds of protein expression plasmids as well as clones that influence and report protein expression (shRNA, microRNA, reporter plasmids). The lab specializes in parallel small scale screening in bacteria, yeast, insect, and mammalian cells to optimize protein quality.  An innovative “purify first” platform tests multiple expression modalities side by side for the most purifiable form of each target protein. Scale-up to tens of liters of culture and hundreds of milligrams of protein is routine.

Clone Optimization 

  • Custom DNA cloning
  • Subcloning
  • Plasmid DNA preparation
  • Site-directed mutagenesis
  • Custom vector construction
  • Gene optimization
  • shRNA/miRNA expression and reporter constructs
  • Consulting

Eukaryotic Expression 

  • Expression of recombinant proteins using the baculovirus expression vector system (BEVS)
  • Monoclonal antibody production
  • Recombinant protein expression in mammalian cells

Microbial Expression

  • Transformations of bacteria and yeast strains
  • Colony screening for expression optimization in yeast
  • Expression screening in bacteria to maximize recombinant protein quantity and quality
  • Batch, batch-fed, or continuous fermentation in multiple yeast and bacteria strains (2ml to 60L)
  • TFF Concentration, Buffer Exchange, and Ultrafiltration
  • Cell lysate preparation and clarification
  • Analysis: SDS-PAGE, Western Blots, Dot Blots, etc.
Protein Purification
  • Micro-scale purification screening and optimization
  • Scale-up from micrograms to grams
  • Endotoxin testing and removal
  • Customized Protein Purification
Mammalian Viruses
 
PEL offers expertise in the production of custom recombinant lentivirus to study transgene or shRNA delivery for gene knockdowns in vitro and in vivo.  Production services are complemented by offering virus concentration and titration services.  Panels of ready-to-ship lentiviruses allow investigators to evaluate different promoters.
 

Contact
Dom Esposito, PhD, Laboratory Director
Tel 301-846-7376
espositod@mail.nih.gov

High-risk projects may be eligible for OSTR subsidies.

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