Contract research

Contract Research

With a foundation of decades of experience in the field of native antigen and recombinant protein contract research, process development and production, NAC is able to undertake custom projects to optimise protocols and manufacture your proteins and antigens. We have 5 development laboratories fully staffed by a team of molecular biologists and development scientists who are able to execute projects as short as a few days through to long term programmes of a year or more.
Our scientific advisory board and academic collaborators, including Oxford University, ensure that we are able to draw upon the latest techniques and thinking which can be brought to bear on your project. Our track record, as exemplified by the case studies below, demonstrates our ability to deliver complex and challenging customer contracts on schedule and within budget.
We are able to offer contract research, contract process development, antigen and recombinant protein production and advisory services in the following areas:

    • DNA design and cloning
    • Vector and/or gene optimisation
    • Recombinant protein expression in mammalian cells, yeast or bacteria
    • Protein purification research and optimisation
    • Native organism growth (virus/bacteria up to BSL 2)
    • Native protein purification
    • Production and purification from established/literature protocols
    • Transfection studies
    • Ultracentrifugation
    • Contract freeze drying / freeze drying optimisation
    • Production of standards and controls

Our core values include openness and honesty. Nothing hampers collaboration more than poor communication and lack of transparency and we have established project and client management systems to ensure that our clients have complete visibility of the status and progress of their project. Our scientists would be delighted to discuss your projects with you and some examples of our recent work are given below:


Contract research case study 1

Dr Holger Schuhmann

Remit: Develop a custom exclusive purification protocol capable of production of 50mg of a pure human intracellular protein for use in immuno and activity assays for a clinical trial. The project was managed by Dr Holger Schuhmann, The Native Antigen Company’s lead purification expert.

Stage 1 – DNA synthesis, cloning, plasmid and protein yield and expression studies (9 weeks)

24 permutations of optimised DNA, bacterial host cell line and vector backbone were assessed for plasmid yield in mini, midi and giga prep sizes. Cryptic bacterial promotors that had a deleterious effect on plasmid yield were identified for many constructs, enabling selection of a lead plasmid candidate in a permissive bacterial strain that showed good yield in gigaprep with a 5x increase in protein expression over initial constructs. SDS-PAGE and Western blots were presented to enable a stop/go decision by the customer, thereby minimising upfront costs and risk.

Stage 2 – Scale-up and purification research (12 weeks)
Purified protein
Lead candidate plasmid was bulk prepped and large-scale transient transfection of human suspension cells was performed to supply bulk raw material for purification research. Lysis methods were investigated and optimised, then a purification method was developed. ELISA and Western blot demonstrated that the histidine tag was occluded in the lead candidate construct; accessible when denatured but not when native during chromatography. Rapid recloning was performed to add a linker sequence into the DNA construct, and bulk preparations were repeated. Binding of the protein was then successful and a purification method resulting in 95%+ pure protein was developed. Activity testing was performed using an outsourced customer assay, all other QC assays were performed in house. Using the resultant yield calculations a price per mg for future production was agreed to enable production of 50mg for the clinical trial.
 
 
Figure to the left: Final purified protein

“Thanks for all the hard work you put in on this difficult protein. We will definitely use you again on our next project”
– Case study 1 customer


Contract research case study 2

Dr Holger Hannemann

Remit: Fee for service to perform a method from a published paper to generate pure bacterial antigen for a clinical trial assay. The project was managed by Dr Holger Hannemann, NAC’s lead scientist.

Extensive literature review was performed to elucidate as much of the method as usual. As is always the case with materials and methods sections, the precise SOP was unclear and so some initial develpment work was required. An initial method was performed in the first week, however the resultant desired antigen (16 kDa) was contaminated with a larger protein of just above 50 kDa.

15kDa protein

Further literature searches produced a suspected candidate for the contaminant protein, thereby enabling NAC scientists to hypothesise from which section of the method it was derived. Several optimisations to the method were set in place and the procedure was repeated. This time the contaminating band was removed, resulting in perfect purity. The yield was also increased by 6.5x, resulting in a successful project delivered on time and to budget.

Figure to the left: Week 1 method (lanes 2 and 4), Week 2 method (lanes 3 and 5)

“Your adaptions look promising. Sorry that we cannot help you with more details. But again thanks for helping us out!!”
– Case study 2 customer

 

 

 


If you need an antigen or protein producing as part of any contract research or process development service, from generating 1mg for academic research to generating 100mg for a clinical trial assay, please do get in touch, we are capable and happy to quote for any project.

Simply summarise your proposed project in an email to contact@thenativeantigencompany.com, or ring +44 (0) 1869 238 067 to discuss your requirements, we are always happy to hear from you.

Contract purification

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