Based on the outstanding expertise and rich experience, Creative Biolabs has specially developed a system approach of TCR generation and optimization for TCR development. To make sure to obtain high quality TCR-modified T cells prior to scientific studies, a serial assay of TCR validation assay are needed. More than that, we can tailor a full range of validation assay strategy for your objective.
The T cell receptor is an ab heterodimeric structure that confers antigen specificity and functional avidity to T cells, in association with the CD3 signaling heterocomplex. Since TCR ab gene sequences can be isolated from tumor-reactive T cell clones, one possible approach to generate tumor antigen-specific T cells for therapy is to transfer these ab sequences to a polyclonal or nonreactive T cell population to redirect their specificity. In order to obtain high quality TCR-modified T cell prior to scientific studies, TCR in vitro assay related to TCR construction are needed. Creative Biolabs offers a series of assays for TCR validation including transgene expression, cytokines release, TCR/CD3ζ stability assay and so on. If you have not found out other services that you want, you can contact us for more information. Our TCR validation services include but are not limited to:
TCR transgene expression assay is a method to evaluate TCR gene transfection efficiency, commonly used in the whole process of TCR development, including post-transfection with TCR gene, cell preparation and clinical treatment period to insure the best outcome. Commonly, we offer the classic measurements including western blot (WB), enzyme-linked immunosorbent assay (ELISA), flow cytometry (FCM), immune-histochemical (IHC) and so on. More than that, we will design an optimized strategy for our clients’ special requirements.
When T lymphocytes encounter antigens via the TCR, information about the quantity and quality of antigen engagement is relayed to the intracellular signal transduction machinery. On the other hand, TCR lacks a significant intracellular domain itself. Therefore, the associated CD3 molecules (CD3ε, CD3γ or CD3δ) which contain intracellular signaling domain couple the TCR/CD3 complex as the downstream signaling machinery. That’s why we should do TCR/CD3 stability assay to make sure the engineered TCR can work during the process of cancer therapy. At Creative Biolabs, the TCR/CD3 complex stability assay can be achieved via immunoprecipitation and immunoblot.
A key feature of the adaptive immune response is the ability of clones of antigen-specific lymphocytes to rapidly proliferate and differentiate into effector cells. So, the lymphocyte culture and proliferation assay is needed to monitor an immune response. CFSE-based flow cytometry combined with Trypan Blue staining is used to analyze cell proliferation and viability respectively.
TCR can specifically recognize peptide antigens presented by APCs. The recognition leads to the activation of T cells and a series of functional outcomes like cytokine production. Therefore, to assay the ability of cytokines releasing is an important part for understanding the functional role and efficiency of TCRs. The basic methods for measurement of T cell immune response include ELISpot, ICS, and cytometry. We will choose the assay strategy according to client’s project.
Mismatched pairs of TCR α and β chains may generate novel TCR pairs of undefined and have potentially self-reactive specificity. We can evaluate TCR pairing and carry out optimized strategies to promote proper pairing by cell-based emulsion RT-PCR, generation of an exclusive TCR heterodimer, murine-human hybrid TCR or chimeric TCR.
Creative Biolabs has the capability to enable you to free up your time for core work and project. Our service can be designed to meet your special needs if you have any requirements. If you are interested in our service, please contact us by E-mail and our team will get back to you as soon as possible.
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