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TCR Pairing

Creative Biolabs provides various techniques such as flow cytometry-based FRET, immunoprecipitation and western blotting for TCR pairing assay. Mispaired TCR heterodimers may result in the diluted expression of TCR transgene or the generation of autoreactive TCR, which affect the efficacy and safety of TCR engineered T cell therapy subsequently. It is a challenge to optimize TCR pairing issue.

TCR pairing

Figure: TCR α- and β-chain pairing and mispairing. 1 and 2: pairing; 3 and 4: mispairing
(Disease Models & Mechanisms, 2015)

In the strategy of TCR-transfer, the modified T cells often have lower avidity than the parental T cells, which is one difficulty to achieve wild-type levels of TCR expression, resulting in limited efficacy. In order to solve the problem, the TCR chains should be initially selected for appropriate affinity and the vectors need to achieve and maintain high-level expression. Even these factors are met, there is also the possibility that the introduced exogenous TCR α and β chains assemble with the exogenous TCR αβ pairs, which decrease T cell avidity. Besides these, mismatched pairs may generate novel TCR pairs of undefined and have potentially self-reactive specificity. Various strategies have researched to facilitate TCR chains matched pairing. It is demonstrated that introducing cysteines residues into the constant region of TCRα and TCRβ chains can promote preferential pairing with each other, increase total surface expression of the introduced TCR chains, and reduce mismatching with endogenous TCR chains.

Moreover, 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. Our customer-oriented services are supplied to meet your requirement about TCR.

In the process to assess TCR pairing, flow cytometry-based FRET, immunoprecipitation and western blotting are generally used. Besides these classical methods, probes-based approach can also be developed. Based on the specific requirements of every client, scientists from Creative Biolabs can design and select appropriate strategies to assess TCR gene transfer.

References:

  1. Michaela Sharpe and Natalie Mount. Genetically modified T cells in cancer therapy: opportunities and challenges. Dis Model Mech. 2015 Apr; 8(4):337-50.
  2. Jürgen Kuball, et al. Facilitating matched pairing and expression of TCR chains introduced into human T cells. Blood. 2007 Mar 15;109(6):2331-8.
  3. Minying Zhang, et al. A new approach to simultaneously quantify both TCR α- and β-chain diversity after adoptive immunotherapy.

To discuss your demands or to request a proposal, please contact us by info@creative-biolabs.com.

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CONTACT US

USA
45-1 Ramsey Road, Shirley, NY 11967, USA
Tel: 1-631-871-5806
Fax: 1-631-207-8356
Email:

Europe
Ringstrasse 4, 64401 Gross-Bieberau, Germany
Tel: 44-207-048-3343

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