scFv Generation

Creative Biolabs offers customers superior approaches for scFv generation such as the one-stop solution for developing CAR from Hybridoma or Phage Display, which manifest as mature and delicate technologies for scFv construction. Particularly, a CAR-T Library Technology on our platform makes great contribution to the selection of desired scFv, whose expression and function are correlated with T cell activation activity.

Basically, a variable region (Fv) consists of a heavy chain variable region (VH) and a light chain variable region (VL), while a single-chain variable fragment (scFv) normally consists of a VH and a VL connected together via a peptide linker. As the most critical initial step, the generation of specific scFv and optimization of its superior affinity lay down the most basic foundation of an effective and specific CAR-T therapy. Without a doubt, a superior scFv leads to an even more superior CAR-T therapy. Creative Biolabs manifests as a world leading expert in antibody engineering field and provides our clients with the best scFv with the most superior affinity in order to meet their research needs. Currently, we offer the two strategies to generate scFv.

A. Generate scFv from Hybridoma Cell Lines as the following procedures:

Figure 1 Flowchart of scFv generation from Hybridoma (Creative Biolabs)

1. Immunize mice with the antigen, and then remove the spleen to get splenocytes (plasma B cells);
2. Fuse the acquired splenocytes with the cultured myeloma cells to establish hybridoma cell lines following by the cell limited dilution and affinity screen;
3. Convert the Fv from the discovered full monoclonal antibody into a scFv, which can target the tumor antigen our customer interested in;
4. Clone the superior scFv with CAR components to construct a Lentivirus vector for the future steps.

B. Generate scFv from Phage Display

At Creative Biolabs, the scFv Phage Display is established originally with an immunized phage display scFv library against the interested antigen. Creative Biolabs guarantees the scFv phage library to achieve a capacity of over 1×10¹⁰ clones. The detailed procedures are depicted as the following:

1. Design the suitable primers for cloning the antibody genes from B lymphocytes of several different species, followed by integrating the antibody genes into a phage display vector;
2. Transform the recombinant phagemid population into Escherichia coli to produce phage-displayed scFv depending on the helper phage;
3. Screen the scFv phage display library with the specific antigen from our customers or the recombinant antigen prepared at Creative Biolabs;
4. Sequence the identified scFv and optimize the affinity with multiple parameters.

Figure 2 Flowchart of scFv generation from Phage Display (Creative Biolabs)

In addition, Creative Biolabs also provides optional molecular characterization service during scFv generation, such as western blot, ELISA or other customized requests. Of note, Our world-leading CAR-T library technology make it possible to screen high-affinity and functional CARs based on T cell display and T cell activation. With the aid of this powerful technology, the selected stable clones can be applied in clinic trails immediately. Several CAR-T libraries against CD19, Her2, Her3, EGFR, FGFR1, VEGFR, etc are available to use.

With professional service and various technology platforms at Creative Biolabs, we offer the best screening and optimization for superior affinity. Meanwhile, the customs sequences of scFv are also welcomed for further optimization, cDNA synthesis, and subsequent service. Our years of experience guarantee we can offer you our high-quality scFv generation. Creative Biolabs offers you world-class scFv construction services to greatly accelerate your research progress with much lower unpredictable risk. Please do not hesitate to contact us.

CAR Biomarker indentification & selection High affinity scFv generation CAR design & construction Virus packaging & CAR transfection CAR in vitro assay CAR Pre-clinical in vivo assay CAR clinical trail