TCR-T Lead Sequence Development Service-Tekbiotech-Yeast Display Service,Phage display technology

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TCR-T Lead Sequence Development Service

With more than ten years of business experience and deep experience in antibody development, TekBiotech's expert team has successfully built an efficient antibody drug candidate molecule platform, which not only covers the development of TCR-T lead sequences, design services for CAR-T and CAR-NK lead antibody sequences, but also widely provides discovery services such as scFv antibodies, VHH antibodies and Fab antibodies, all of which show excellent quality. Relying on advanced phage display technology and single B cell sequencing technology, TekBiotech has made great achievements in the field of antibody development and can quickly provide customers with TCR-T customized development and CAR antibody lead sequence development services.

In the development of TCR-T lead sequences, the correct pairing of TCR-α chain and TCR-β chain is crucial. TechBio uses professional tools to optimize it and can select suitable vectors to optimize its expression conditions. Based on the next-generation sequencing technology, TekBiotech can identify and locate target antigens and sort out specific CD8+T cells by flow cytometry. In addition, through single-cell sequencing technology, TekBiotech can separate tumor-specific TCR encoding genes with high throughput. TekBiotech can insert TCR gene fragments into the phage coat protein (such as pIII) sequence, so that the TCR sequence can be displayed on the phage surface. The constructed phage library can reach 10^8-10^9, ensuring that the library diversity, insertion rate and positive rate exceed 90% to meet the quality requirements of different customers for antibody display libraries.

At the same time, TekBiotech provides comprehensive antibody development downstream services, including the preparation and amplification of TCR-T cells in TCR-T cell therapy, cytokine release testing, in vitro cytotoxicity and proliferation testing, cell viability and biodistribution analysis, etc., to ensure the safety and effectiveness of TCR-T cells, as well as one-stop technical services such as cell killing verification. TekBiotech scientists can analyze and design reasonable solutions based on the specific experimental needs of customers, aiming to provide customers with personalized customized solutions to accurately meet their needs.

█ TCR-T Lead Sequence Development Service

In addition to membrane proteins, proteins in cells can also be recognized by TCR-T cells, which shows that it targets a wider range of targets. And TCR-T can be activated at a lower epitope density. This characteristic also improves the detection and clearance capabilities of TCR-T cell therapy. Generally speaking, its affinity is lower than that of CAR, which means that in TCR-T cell therapy, each TCR-T cell may be allowed to detect and eliminate several antigen-presenting tumor cells. TekBiotech's TCR-T customized development service based on phage display technology uses phage display libraries to obtain corresponding sequences, which can enable cells to have corresponding clearance capabilities. The specific process is shown in Figure 1.

TCR-T Lead Sequence Development Service-tekbiotech1.png

Figure1 TCR-T Custom Development Service Flowchart Based on Phage Display Technology

█ Service Content and Cycle

Steps	Service Content	Period
Step 1: Animal Immunization	(1) Animals are immunized 4 times, with one booster shot, for a total of 5 shots; (2) Collect negative serum before immunization, and collect blood for the 4th shot for ELISA to test serum titer; (3) If the 4th shot serum antibody titer meets the requirements, another booster shot is performed 7 days before blood collection. If it does not meet the requirements, continue routine immunization; (4) If the titer is qualified, blood is collected to isolate T cells;	10 Weeks
Step 2: cDNA Preparation	(1) Extraction of total RNA from T cells (RNA extraction kit); (2) Preparation of cDNA by high-fidelity RT-PCR (reverse transcription kit);	1 Working Day
Step 3: Construction of Antibody Library	Based on phage display technology: (1) Using cDNA as a template, two rounds of PCR are used to amplify the genes of the α and β chains of TCR; (2) Phagemid construction and transformation: TCR gene fragments are spliced into phagemid vectors, electroporated to transform TG1 host bacteria, and antibody library is constructed; (3) Identification: 24 clones are randomly selected, and PCR is used to identify the positive rate + insertion rate; (4) Auxiliary phage preparation: M13 phage amplification + purification; (5) Antibody display library rescue;	3-4 Weeks
Step 4: Antibody Library Screening (3 rounds)	Based on phage display technology: (1) Default 3 rounds of screening (solid phase screening): pressure screening to remove non-specific antibodies to the maximum extent; (2) Pick a single clone to amplify phage + IPTG induced expression + ELISA to detect positive clones; (3) Pick all positive clones for gene sequencing;	4-5 Weeks