Cancer Sciences Protein Facility

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About the Cancer Sciences Protein Facility

The Cancer Sciences Protein Facility has all the equipment and expertise to produce tailored recombinant proteins.  

Our focus is primarily on proteins and antibodies involved in cancer, but we also produce proteins for other applications. 

The Protein Facility has produced over 100 different proteins since 2006, contributing to over 45 articles. The protein production incorporates the whole process from cloning and expression to purification and quality control. 


Our highlights include:

  • Cloning of the gene of interest in expression vector 
  • Overexpression of proteins in bacteria, insect cells or mammalian cells 
  • Purification of protein by affinity, ion exchange and size exclusion chromatography 
  • Applications: Cell culture, flow cytometry, microscopy, ELISA, crystallography 

Technical specification

Molecular biology

  • Cloning with standard restriction digestion and ligation techniques 
  • Cloning with sequence ligation-independent cloning (SLIC) techniques 
  • Design of synthetic DNA with sequence optimisation; PCR, Inverse-PCR, mutagenesis 
  • Making of high competent bacteria for cloning (JM109 and others) 
  • Bacterial vectors: pET (His6), pGEX (GST), pOPIN (His6, GST, MBP, SUMO, TG, NusA, Thioredoxin) 
  • Insect vectors: pMT-Puro, pFastBac, pOPIN 
  • Mammalian vectors: pcDNA3, pDSG (His6, TwinStrep), pCI-Puro, pEE6.4

Expression systems


Pros: High levels of protein, quick, low cost, scalable, simple culture conditions.

Cons: Difficult with large proteins, proteins can be misfolded, lack of post-translational modifications.

  • E. coli BL21 Strains DE3, pLys, pLacI, Rosetta2 
  • Culture in LB and Terrific Broth media 
  • IPTG induction and auto-induction

Insect cells  

Pros: Closer to mammalian protein processing.

Cons: More demanding culture conditions than bacteria, production of baculovirus can be time consuming.

  • Stable transfection of S2 cells with copper-inducible system 
  • Baculovirus system with Sf9 and High5 cells

Mammalian cells  

Pros: Natural folding, post-translational modifications, pyrogen-free

Cons: Expensive 

  • HEK MEXi293E cells - transient expression 
  • ExpiCHO cells - transient expression  

Protein purification

  • Refolding of insoluble proteins expressed in bacteria 
  • Affinity chromatography for tagged proteins (His6, GST, MBP, TwinStrep), antibodies and Fabs 
  • Ion exchange chromatography
  • Size exclusion chromatography

Protein modification

  • Tag removal: TEV, H3C, SUMO proteases 
  • Labelling of proteins and antibodies  
  • Endotoxin removal from protein sample

Specific equipment

  • MaxQ 6000 Shaker incubators for bacterial expression (room temperature – 37C) 
  • MaxQ 6000R Refrigerated shaker incubator for bacterial expression (4C – 37C) 
  • Infors HT Multitron shaker incubators for mammalian transient expression 
  • Infors HT Minitron shaker incubator for insect expression 
  • Cytiva Akta PrimePlus protein purification systems 
  • Bio-Rad NGC Protein purification system 
  • Amicon stirred cell concentrator


  • Flow cytometry 
  • Fluorescent microscopy 
  • Protein-protein interaction – BiaCore, ELISA 
  • Enzymatic assays 
  • Cell culture 
  • Mouse work 
  • Protein Structure in collaboration with IfLS groups

Examples of recombinant proteins produced by the Protein Facility

  • Tag removal: His6-H3C / GST-H3C, His6-TEV and SUMO proteases  
  • Biotinylation ligase Bir A enzyme 
  • Annexin V-FITC and Annexin V-APC  
  • MHC I tetramers  
  • Cytokines and interleukins 
  • Antibodies, Fabs and ScFv 
  • Tailored proteins 


Contact us

We welcome your enquiries.
Centre for Cancer Immunology Room 3049Coxford RoadSouthampton General HospitalCentre for Cancer Immunology, MP127Southampton SO16 6YD