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CRB Discovery
  In this issue
  Case Study:
Analysis of the Molecular Nature of CTLA-4 using Custom
Monoclonal and Polyclonal Antibodies
INFINITY™ antibodies:
The advantages of a CRB monoclonal antibody
Key to our Success:
Expert project design and optimised strategies
  Analysis of the Molecular Nature of CTLA-4 using Custom Monoclonal and Polyclonal Antibodies  
Characteristics of the CTLA-4 gene, required for immune homeostasis, were investigated by the Wicker group at the Cambridge Institute for Medical Research (CIMR) who commissioned the preparation of peptide-induced custom monoclonal and polyclonal antibodies specific for the soluble protein isoform of CTLA-4, sCTLA-4, from Cambridge Research Biochemicals.

The innovative peptide design by Cambridge Research Biochemicals allowed the group to obtain antibodies that distinguish sCTLA-4 from the transmembrane isoform, Tm-CTLA-4.

Using these antibodies, Laura Esposito in the Wicker group has demonstrated that the sCTLA-4 protein is secreted at low levels following the activation of primary human CD4+ T cells.

With sCTLA-4-specific antibodies in hand to differentiate the two isoforms of CTLA-4, the investigators were able to uncover the involvement of Tm-CTLA-4 associated with microvesicles produced by activated human CD4+ T cells. The group continues to elucidate the functional roles of sCTLA-4 and microvesicle-associated Tm-CTLA-4 through subsequent work.

The 4B8 mAb,10D1 mAb, and 4017 polyclonal Ab validation by Western blot under reducing conditions on detergent lysates of HeLa cells expressing human recombinant Tm-CTLA-4, sCTLA-4, and vector only. Specific detection of Tm-CTLA-4 is performed with CTLA-4 (C19) polyclonal Ab.

This figure was first published in the Journal of Immunology (Esposito et al, 2014,
With thanks to Professor Linda Wicker (Cambridge Institute for Medical Research) for granting permission to highlight her work.
Linda Wicker  
Linda Wicker
Group Leader at the Cambridge Institute
for Medical Research
CIMR Wellcome Logo

“I am very impressed with how well the process of generating custom antibodies with the Team at Cambridge Research Biochemicals worked, particularly as it was extremely important for our research to obtain isoform-specific monoclonal antibodies. The Team at Cambridge Research Biochemicals were knowledgeable and collaborative throughout the process, engaging in much discussion to ensure that we received high quality, fit-for-purpose antibodies. I have recommended to my colleagues that they consider CRB for their antibody needs”.
INFINITY Antibodies
Cambridge Research Biochemicals (CRB) has provided a top quality custom monoclonal antibody production service to the life science community for over eight years.

A recent expansion and investment programme has permitted the development of a brand new cell culture suite on site. This has allowed CRB to substantially increase the capacity for monoclonal antibody production which will lead to a faster service for our customers.

Monoclonals offer many advantages over traditional polyclonals:

  • The antibody generated is available in perpertuity due to the immortal nature of the cell line.
  • Monoclonal antibodies have a defined specificity and a low non-specific activity.
  • They are ideal for low abundance antibodies, such as IgM.
  • Positive & negative screens can be peformed to improve the specificity of the antibody.
  • Multiple cell lines can be purchased – the additional clones/cell lines generated can be purchased by the customer.
  • Monoclonal antibodies offer reproducibility and traceability.

At Cambridge Research Biochemicals, we offer a comprehensive Monoclonal antibody service which follows our successful four-phase project outline:

  • Expert advice on the vital first step of antigen selection from either your target protein or peptide sequence
  • Design of screening strategy to select for clones against specific PTMs or protein regions
  • Choice of appropriate species (mouse, rat, rabbit)

  • Extended immunisations to achieve optimum titre prior to cloning
  • Development of screening assay to identify the best clones
  • Multiple sub-cloning to capture stable cell lines
  • Scale-able production and purification capacity

  • Regular progress updates and Technical assistance
  • Defined check points for project progression
  • Written reports available at end of each phase
  • Back-up freezing of sub-clones and final cell line

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