Considerations in CAR Lead Discovery

CAR-T cell therapy has several limitations, including CAR-T cell-associated toxicities, on-target/off-target effects, antigen escape, undesirable trafficking & tumor infiltration, and immunosuppressive microenvironment, etc.. CARs which serve as the targeting component, consist of an extracellular binding domain, a hinge domain, a transmembrane domain, and one or more intracellular domains, all of which influence the efficacy and safety of CAR-T cell therapy. When designing CAR molecules, various factors typically need to be taken into consideration.

Considerations in CAR Lead Discovery

CAR Lead Discovery Strategy

GenScript ProBio created a comprehensive antibody discovery solution tailored for CARs, incorporating five distinct technical campaigns to yield diverse binders. Our offering sophisticated in vitro and in vivo bioassays, along with an industry-leading lead optimization platform. These tools swiftly identify CARs with robust specificity, suitable affinity, and impressive diversity, empowering you to capitalize on market opportunities quickly.

  • Lead Generation

    Lead Generation

    • Hybridoma discovery
    • Single B cell screening
    • Human naïve library
    • sdAb immunized library
    • sdAb naïve library
  • Lead Optimization

    Lead Optimization

    • Antibody humanization
    • Antibody affinity maturation
  • In Vitro Assay

    In Vitro Assay

    • CAR-T cell generation
    • In vitro functional assay
  • In Vivo Assay

    In Vivo Assay

    • Efficacy
    • PK study
    • Toxicity
Discovery Strategy for Therapeutic Antibody VS CAR Lead
    • The evaluation standard of a therapeutic antibody is first its biological function and then affinity etc..
    • So functional assays are conducted right after the primary screening
    • The screening scale is larger to increase the chance of getting leads with good function.
    • ScFv, but not mAb, is expressed in CAR lead generation.
    • Because the role of an antibody in a CAR-T development is mainly a targeting tool, affinity is the top consideration when selecting scFv or sdAb candidates.
    • Specificity test can be performed according to customer requirements.
    • The function of CAR-T cells not the scFv or sdAb itself should be validated in a CAR-T development project.

CAR Lead Discovery Service

CAR lead candidates typically use sdAbs or scFvs as the ideal structural components, which need a suitable affinity for precise targeting while minimizing the risk of cellular toxicity. Furthermore, it's essential to consider the other attributes, such as antibody cross-reactivity, developability, stability, and candidate differentiation.

GenScript ProBio offers tailored antibody discovery solutions for CAR lead candidates, presenting five distinct technical campaigns for sdAbs and scFvs. Following initial screening, we will directly perform SPR affinity ranking to yield an array of binders with varied affinities. Through additional functional assays like epitope binning, cross-reactivity testing, and early developability testing, our services expedite the acquisition of high-quality CAR lead candidates with unique properties.

  • Proven Excellence

    750+ projects completed

    14 projects advanced to clinical trials

    • Elimination of cumbersome subcloning steps for greater efficiency
    • Achieve purified antibody within an impressive 2 months (standard immunization)
    • Conducting high-throughput assays with hybridoma supernatant for rapid, reliable results
  • Extremely Fast, Minimal Diversity Loss

    100+ projects completed

    • As fast as 1.5 months to get purified fully human antibodies combining transgenic animals with single B cell screening platform
  • High capacity Exceeding 10^11, High diversity, High quality

    • Obtain fully human antibodies as fast as 1 month
    • The high-quality naïve library increases the probability of screening the high-potential Ab leads
    • The diverse panning and screening strategy ensures affinity and diversity of Ab leads

By 2023.11

1 FDA-approved drug6 global licensing deal; 150+ sdAb lead generation projects

  • sdAb Naïve Library Service
    • 300 Alpaca donors, Library size: 2×1011
    • In frame rate/ORF rate: >95%
    • High diversity
    • Get purified antibodies as fast as 1.5 months
  • sdAb Immunized Library Service
    • Easy access to Alpaca breeding farm
    • Using naïve alpaca for each project
    • High affinity
    • Success rate: 89%

By 2023.11

CAR Lead Optimization Service

GenScript ProBio has a comprehensive antibody engineering platform (antibody humanization, affinity maturation, developability optimization, Fc engineering) to optimize your antibody candidates:

In Vitro Functional Validation

ProBio's comprehensive CAR-T/NK in vitro pharmacology platform offers a range of services to support you in developing CAR cell therapy. Our services include the generation of CAR encoding vector, T cell activation and transduction, expansion of CAR-T cells, and functional assay for cytotoxicity and cytokine release.

Generation of CAR
encoding vector
T cell activation and
Expansion of CAR-T
Function assay for cytotoxicity
and cytokine release
    • FTO CAR vector
    • Non-virus transfection
    • Lentivirus
    • Retroviral
    • High fold expansion
    • Time-based killing assay
    • Cancer cell lines
    • Overexpression cell lines
    • Luciferase Target cells

CAR-T functional evaluation

CAR-T functional evaluation

CAR-T In Vivo Pharmacology

The efficacy and safety of CAR-T need to be simultaneously verified in early pharmacology studies. Genscript ProBio provides a one-stop CAR-T in vivo pharmacology solution throughout the whole discovery process from drug design and screening to PCC confirmation, which will help you obtain high-potential CARs for optimal therapeutic outcomes.

Key Material Screening &
Method Development
CAR-T Manufacture
In Vivo Efficacy
In Vivo PK Package
Ex & In Vivo Toxicity

CAR-T functional evaluation

CAR-T functional evaluation
  • Figure A:Solid Tumor(MDA-MB-231),Single Dose CAR-T Treatment inhibit In Vivo Tumor Growth

    Figure A:Solid Tumor(MDA-MB-231),Single Dose CAR-T Treatment inhibit In Vivo Tumor Growth

  • Figure B:Single Dose CAR-T did not influence Mice Body Weight and Temperature (Data not show)

    Figure B:Single Dose CAR-T did not influence Mice Body Weight and Temperature (Data not show)

  • The quantities of circulating CART Cells

    Figure C:Circulating CAR-T Cells in peripheral blood works as a three-phase kinetics diagram*

    Figure C:Circulating CAR-T Cells in peripheral blood works as a three-phase kinetics diagram*

  • Figure D:CAR Positive T Cells can accumulate In tumor

    Figure D:CAR Positive T Cells can accumulate In tumor

Case Study

  • Feel free to download the poster to discover how we leverage the naïve VHH phage display library and an integrated antibody engineering platform to create antibody leads for CAR-T therapy. In this case study, an alpaca naïve VHH library was panned and screened by ROR1 proteins of 3 different ECDs and CHO-K1/ROR1 cell line to identify ROR1 VHH candidates. These VHH candidates were used to construct CAR-T cells and further evaluated by vitro functional assays, based on which final leads were selected for optimization by affinity maturation and ready for the development of CAR-T therapies.

    Watch Our On-demand Webinar

    Utilization of VHH phage display library & integrated protein engineering platform to generate antibody leads for CAR-T therapy

  • GenScript ProBio_generate antibody leads for CAR-T therapy