Protein Expression Facility (PEF)

The Protein Expression Facility (PEF) provides a recombinant expression and purification service to the University of Birmingham Research community.

As part of the Technology hub the PEF acts as a focal point for advice about all aspects of recombinant protein production. The expression systems used by the PEF will enable the recombinant proteins generated to be used in a wide range of downstream applications.

Protein Expression Facility Team

Protein Expression Specialist

Jamie R. M. Webster

Jamie joined the Protein Expression facility in 2013. He was awarded a BSc (Hons) in Applied Science (1986) from the Polytechnic, Wolverhampton, a MSc in Science (F12 - frontiers in medical science) with Distinction from the Open University (2009) and holds a current RSci from the Science Council (IST 2015-2020, RSB 2020 to date). Jamie has thirty-six years technical experience both with the civil service (MAFF), NHS (NBTS) and HEI’s (Cambridge, Nottingham and Birmingham). His experience prior to joining the facility supported research in Cyanobacterial proteins and Photosystem II, Archaeal Cas3 and Cas5, the Chromosomal passenger complex and Wallerian Degeneration and since joining the facility has supported colleagues in the college of MDS, LES and School of Chemistry.

Email:  J.R.M.Webster@bham.ac.uk

Research Facilities Manager

Dr Adriana Flores-Langarica

Email: A.FloresLangarica@bham.ac.uk

Protein Expression Academic Lead

Professor Benjamin Willcox

Email: b.willcox@bham.ac.uk

Why work with us?

Equipment in the Protein Expression Facility (PEF)

The PEF provides a service as close to cost price as possible, but consistent with facility sustainability. The PEF aims to cater for the production of a wide variety of proteins to enable a comprehensive range of downstream technologies and to be responsive to customer needs where possible.

The PEF reside within a designated, fully-equipped laboratory suite that was funded by a central infrastructure bid in 2010 through the College of Medical and Dental Sciences.

The PEF is self-funded, with annual running costs met through recharging schemes proportional to the level and nature of collaboration. Activities are distributed across the Birmingham campus research community, but the PEF welcomes collaboration with Midlands Innovation and external companies.

The PEF provides a central location to undertake recombinant protein expression within a dedicated environment. It also provides a range of expression and purification services or training which can be commissioned at any time.

Who do we work with?

The Protein Expression Facility (PEF) at the University of Birmingham is available to the following groups:

  • Current staff and students
  • Prospective staff and students
  • Academics both internal and external
  • Industry professionals

For more information regarding how to get in touch with the PEF, please use the contact details at the bottom of this webpage.

Resources and Services

The PEF offers a wide range of purification techniques, expression models and access to equipment. Examples include:

PEF - Resources and Services image 1
  • Expression construct design and bioinformatics
  • Dedicated culture facilities for prokaryotic and eukaryotic expression
  • Expression systems available: E.coli, mammalian cell and insect cell expression
  • Fast Protein Liquid Chromatography
  • PHERAstar FS microtiter plate reader
  • Affinity purification
  • MHC Class I Monomer production
  • Training in purification techniques
  • Endotoxin testing

 

Expression and Purification Facility

PEF - Resources and Services image 2

The PEF offers a one stop shop for small scale test expressions through to medium scale expressions followed by purification and polishing of the protein requested.

The service provided may be inclusive of cloning, optimisation of the construct via test expression through to scaling up for purification and polishing, or may be a Principal Investigators well characterised protocol. Work undertaken will depend on the initial discussions to outline the project requirements and the status of existing Biosafety approvals.

The PEF offers expertise in E.coli, mammalian cell and insect cell expression and has two dedicated culture facilities. One is dedicated to prokaryotic work and has three programmable incubators capable of growth conditions between 4-80 degrees with two dedicated Beckman Avanti centrifuges. The eukaryotic culture facility hosts incubators for static, shaking and roller bottle culture of mammalian or insect cell. 

Downstream applications of refolded MHC1 Protein table, Top to bottom: Crystallographic Structure Determination, Surface Plasmon Resonance Studies, Circular dichroism (CD) measurements and flow cytometry

MHC class I refolds

Biotinylated MHC Class I Monomer are composed of a particular peptide in combination with a particular MHC class I allele that has been biotinylated at the C-terminus of the heavy chain extracellular domain. The Biotinylated Class I MHC/peptide complex can be used in research studies using the MHC tetramer assay and the PEF has been routinely supplying the Birmingham Researcher with monomers since 2010.

The PEF routinely makes HLA A0101, HLA A0201, HLA B8 and HLA B7 and holds a supply of the inclusion bodies used to refold the peptides chosen by the customer.

For more information regarding how to get in touch with the PEF about Biotinylated MHC Class I Monomer production, please use the contact details at the bottom of this webpage.

Surface Plasmon Resonance - Biacore T200

Surface Plasmon Resonance - Biacore T200

Access to a Biacore T200 is available for internal and external customers to use with booking via ppms. The equipment is run as a user operated service and training can be arranged for researchers. For more information regarding how to get in touch with the PEF about the Biacore T200 and for any training requests, please use the contact details at the bottom of this webpage.

Current Research

Currently the facility is collaborating across campus on a range of expression projects with Institutes in the Medical School, Biosciences, Chemistry and with the Midlands Innovation group of Universities.

The PHERAstar provides collaborative resources for the Birmingham Researcher wishing to read single plate fluorescent or chemiluminescent assay’s and our customer range from Sports and exercise to the Medical school. Initial training for the PHERAstar is available from the Canvas course Technology Hub Online Training Resource.

Recent Publications

Eldershaw, S.A., Pearce, H., Inman, C.F., Piper, K.P., Abbotts, B., Stephens, C., Nicol, S., Croft, W., Powell, R., Begum, J., Taylor, G., Nunnick, J., Walsh, D., Sirovica, M., Saddique, S., Nagra, S., Ferguson, P., Moss, P. and Malladi, R. (2021), DNA and modified vaccinia Ankara prime–boost vaccination generates strong CD8+ T cell responses against minor histocompatibility antigen HA-1. Br J Haematol. https://doi.org/10.1111/bjh.17495

Odintsova, E., Mohammed, F., Trieber, C. et al. Binding of the periplakin linker requires vimentin acidic residues D176 and E187. Commun Biol 3, 83 (2020). https://doi.org/10.1038/s42003-020-0810-y

Pedrera M, Macchi F, McLean RK, Franceschi V, Thakur N, Russo L, Medfai L, Todd S, Tchilian EZ, Audonnet JC, Chappell K, Isaacs A, Watterson D, Young PR, Marsh GA, Bailey D, Graham SP, Donofrio G. Bovine Herpesvirus-4-Vectored Delivery of Nipah Virus Glycoproteins Enhances T Cell Immunogenicity in Pigs. Vaccines (Basel). 2020 Mar 2;8(1). pii: E115. doi: 10.3390/vaccines8010115.

Mohindar M.Karunakaran, Carrie R.Willcox, Mahboob Salim, Daniel Paletta, Alina S.Fichtner, Angela Noll, Lisa Starick, Anna Nöhren, Charlotte R.Begley, Katie A.Berwick, Raphaël A.G.Chaleil, Vincent Pitard, Julie Déchanet-Merville, Paul A.Bates, Brigitte Kimmel, Timothy.J.Knowles Volker Kunzmann, Lutz Walter and Thomas Herrmann. Butyrophilin-2A1 Directly Binds Germline-Encoded Regions of the Vγ9Vδ2 TCR and Is Essential for Phosphoantigen Sensing Immunity Volume 52, Issue 3, 17 March 2020, Pages 487-498.e6 https://doi.org/10.1016/j.immuni.2020.02.014

Fiyaz Mohammed, Daniel H. Stones Benjamin E. Willcox, Application of the immunoregulatory receptor LILRB1 as a crystallisation chaperone for human class I MHC complexes, Journal of Immunological Methods, Volume 464, January 2019, Pages 47-56, https://doi.org/10.1016/j.jim.2018.10.011

Jamshad M, Knowles TJ, White SA, Ward DG, Mohammed F, Rahman KF, Wynne M, Hughes G1, Kramer G, Bukau B, Huber D. The C-terminal tail of the bacterial translocation ATPase SecA modulates its activity, Elife. 2019 Jun 27;8. pii: e48385. doi: 10.7554/eLife.48385

Willcox CR, Vantourout P, Salim M, Zlatareva I, Melandri D, Zanardo L, George R, Kjaer 4, Jeeves M, Mohammed F, Hayday AC, Willcox BE. Butyrophilin-like 3 Directly Binds a Human Vγ4+ T Cell Receptor Using a Modality Distinct from Clonally-Restricted Antigen. Immunity. 2019 Nov 19;51(5):813-825.e4. doi: 10.1016/j.immuni.2019.09.006. Epub 2019 Oct 15.

Davey, Martin S.; Willcox, Carrie R.; Hunter, Stuart; et al (2018) The human V delta 2(+) T-cell compartment comprises distinct innate-like V gamma 9(+) and adaptive V gamma 9(-) subsets .NATURE COMMUNICATIONS Open access indicator Volume: 9 Article Number: 1760 Published: MAY 2 2018

Hunter, S., Willcox, C., Davey, M., Kasatskaya, S., Jeffery, H., Chudakov, D., Oo, Y., Willcox, B., Human liver infiltrating γ δ T cells are composed of clonally expanded circulating and tissue-resident populations, Journal of Hepatology (2018), doi: https://doi.org/10.1016/j.jhep.2018.05.007

Salim M, Knowles TJ, Baker AT, Davey MS, Jeeves M, Sridhar P, Wilkie J, Willcox CR, Kadri H, Taher TE, Vantourout P, Hayday A, Mehellou Y, Mohammed F, Willcox BE. (2017) BTN3A1 Discriminates γδ T Cell Phosphoantigens from Nonantigenic Small Molecules via a Conformational Sensor in Its B30.2 Domain. ACS Chem Biol. 2017 Sep 14. doi: 10.1021/acschembio.7b00694. [Epub ahead of print]

K A Khan, A J Naylor, A Khan, P J Noy, M Mambretti, P Lodhia, J Athwal, A Korzystka, C D Buckley, B E Willcox, F Mohammed and R Bicknell Multimerin-2 is a ligand for group 14 family C-type lectins CLEC14A, CD93 and CD248 spanning the endothelial pericyte interface Received 13 September 2016; Revised 6 April 2017; Accepted 14 April 2017 Advance online publication 3 July 2017 oncogene advance online publication 3 July 2017;doi: 10.1038/onc.2017.214

AlAmri MA, Kadri H, Alderwick LJ, Simpkins NS, Mehellou Y. Rafoxanide and Closantel Inhibit SPAK and OSR1 Kinases by Binding to a Highly Conserved Allosteric Site on Their C-terminal Domains.ChemMedChem. 2017 May 9;12(9):639-645. doi: 10.1002/cmdc.201700077. Epub 2017 Apr 12.

Mohammed F, Stones DH, Zarling AL, Willcox CR, Shabanowitz J, Cummings KL, Hunt DF3, Cobbold M, Engelhard VH, Willcox BE.(2017) The antigenic identity of human class I MHC phosphopeptides is critically dependent upon phosphorylation status. Oncotarget. 2017 Apr 8. doi: 10.18632/oncotarget.16952. [Epub ahead of print]

Davey, MS ; Willcox, CR ; Joyce, SP; Ladell, K; Kasatskaya, SA  ; McLaren, JE  ; Hunter, S  ; Salim, M ; Mohammed, ; Price, DA Chudakov, DM ; Willcox, BE Clonal selection in the human V delta 1 T cell repertoire indicates gamma delta TCR-dependent adaptive immune surveillance NATURE COMMUNICATIONS Volume: 8 Article Number: 14760 DOI: 10.1038/ncomms14760 Published: MAR 1 2017

Kadri H, Alamri MA, Navratilova IH, Alderwick LJ, Simpkins NS, Mehellou Y. Towards the Development of Small-Molecule MO25 Binders as Potential Indirect SPAK/OSR1 Kinase Inhibitors.Chembiochem. 2017 Mar 2;18(5):460-465. doi: 10.1002/cbic.201600620. Epub 2017 Jan 30.

J. Zuo, C. R. Willcox, F. Mohammed, M. Davey, S. Hunter, K. Khan, A. Antoun,P. Katakia, J. Croudace, C. Inman, H. Parry, D. Briggs, R. Malladi, B. E. Willcox, P. Moss, A disease-linked ULBP6 polymorphism inhibits NKG2D-mediated target cell killing by enhancing the stability of NKG2D ligand binding. Sci. Signal. 10, eaai8904 (2017).

Salim M, Knowles TJ, Hart R, Mohammed F, Woodward MJ, Willcox CR, Overduin M, Hayday AC, Willcox BE. (2016) Characterization of a Putative Receptor Binding Surface onSkint-1, a Critical Determinant of Dendritic Epidermal T Cell Selection. J Biol Chem. 2016 Apr 22; 291(17):9310-21.

Mackley EC, Houston S, Marriott CL, Halford EE, Lucas B, Cerovic V, Filbey KJ, Maizels RM, Hepworth MR, Sonnenberg GF, Milling S, Withers DR. (2015) CCR7-dependent trafficking of RORγ⁺ ILCs creates a unique microenvironment within mucosal draining lymph nodes. Nat Commun. 2015 Jan 9;6:5862.

Marriott CL, Carlesso G, Herbst R, Withers DR. (2015) ICOS is required for the generation of both central and effector CD4(+) memory T-cell populations following acute bacterial infection.. 2015 Jun;45(6):1706-15.

Navarro-Núñez L, Pollitt AY, Lowe K, Latif A, Nash GB, Watson SP. (2015) Platelet adhesion to podoplanin under flow is mediated by the receptor CLEC-2 and stabilised by Src/Syk-dependent platelet signalling. Thromb Haemost. 2015 May;113(5):1109-20.

Pollitt AY, Poulter NS, Gitz E, Navarro-Nuñez L, Wang YJ, Hughes CE, Thomas SG, Nieswandt B, Douglas MR, Owen DM, Jackson DG, Dustin ML, Watson SP. (2014) Syk and Src family kinases regulate C-type lectin receptor 2 (CLEC-2)-mediated clustering of podoplanin and platelet adhesion to lymphatic endothelial cells. J Biol Chem.  2014 Dec 26;289(52):35695-710.

Marriott CL, Mackley EC, Ferreira C, Veldhoen M, Yagita H, Withers DR. (2014) OX40 controls effector CD4+ T-cell expansion, not follicular T helper cell generation in acute Listeria infection. Eur J Immunol. 2014 Aug;44(8):2437-47.

Borgognone A, Navarro-Núñez L, Correia JN, Pollitt AY, Thomas SG, Eble JA, Pulcinelli FM, Madhani M, Watson SP. (2014) CLEC-2-dependent activation of mouse platelets is weakly inhibited by cAMP but not by cGMP. J Thromb Haemost. 2014 Apr;12(4):550-9.

Contact Us

Requests for quotes and all general enquiries about the PEF, should be directed to:

Telephone: +44 (0)121 414 9266

Email: pef@contacts.bham.ac.uk