scFv Generation and Optimization

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

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.