scFv Generation and Optimization

As the most critical initial step, the production of specific scFv and the optimization of its superior affinity lay the most basic foundation for effective and specific CAR-T/NK/MA therapy. There is no doubt that high-quality scFv can lead to superior CAR cell therapy. Creative Biolabs combines our highly trusted hybridoma technology with our top antibody sequencing and recombinant antibody expression platforms to generate highly specific ScFvs to meet our clients’ needs.

scFv Generated from Hybridoma Cells

The mice were immunized with the antigen, and then the spleen was taken out to obtain spleen cells (plasma B cells).
Spleen cells obtained by fusion of cultured myeloma cells, hybridoma cell lines were established by limiting cell dilution and affinity screening.
Convert Fv from a fully monoclonal antibody found to scFv to target tumor antigens of interest to our clients.
The high quality scFv was cloned with the CAR component to construct a lentiviral vector for future steps.

scFv Generated from Phage Display Library

Design appropriate primers for cloning antibody genes from B lymphocytes of several different species, then integrate the antibody gene into a phage display vector.
Transform recombinant phagemid into E. coli, and produce phage-displayed scFv based on helper phage.
Screening of scFv phage display libraries using our client's specific antigens or recombinant antigen prepared by Creative Biolabs.
The identified scFvs are sorted and the affinities are optimized using multiple parameters.

scFv Generated from Antibody Sequence

Antibody ScFv production from existing antibody sequences is the fastest way to obtain engineered antibody fragments. Our experts can synthesize two variable region gene sequences in vitro and fuse these two regions with the short peptide linker gene (10-25 amino acids). Subsequently, we inserted this long fragment into the vector and expressed it in E. coli or mammalian cell lines (depending on your choice). We also provide further services for purification of this recombinant antibody, making it available for further research.

ScFv Optimization

Efficient T cell activation may vary depending on the target antigen and CAR construct, CAR expression, scFv affinity, spatial restriction, and tumor cell expression of adhesion and costimulatory ligands. scFv can affect CAR function, not just tumor specificity. Creative Biolabs’s researchers can regulate the safety and/or efficacy of CAR T cells through all aspects of scFv design:

For more details, please feel free to contact us for project quotations and more detailed information.