Selection of Antibody Fragments scFv and VHH for CAR-T Cell Therapy from Phage Display Libraries

CAR-T cell therapy emerged as a great promise to cancer treatment. New chimeric antigen receptors (CAR) are needed to be designed and functionally tested to improve the breadth of its use. This chimeric protein CAR associates the binding to a specific antigen to activation signals in the cytoplasm of the T cell, triggering an effector response against the antigen-bearing cell. The crucial modular for CAR structure is the extracellular target-binding domain. This binding domain is generally an antibody fragment, such as scFv (single chain variable fragment), or VHH (a single-domain antibody, or nanobody), that recognize a membrane protein associated to a cancer cell. Selecting a high affinity antibody or antibody fragment to a specific cancer antigen is one of the keys to an effective CAR-T system.

Fusion Biolabs have powerful phage display libraries for this purpose. We have been used phage display technology to isolate antibody coding sequences against extracellular binding domains of tumor associated antigen (TAA) of targeted cancer cell to be used in construction of CAR expression vectors.

The display of antibody libraries on phage is a powerful technique to isolate antibodies for cancer antigens. The most common choice is the construction of high diversity naïve or synthetic libraries that recognize human cancer antigen. The success of the selection is based on two important steps: the sources of high diversity phage libraries, and the selection procedure, which can improve specificity and favor selection of higher affinity mAbs for CAR construction.

Because of this specific antigen, tumor associated antigen (TAA) or cell surface proteins, Fusion Biolabs scientists have developed a specific panning platform for selecting antibody against cell surface proteins: panning in solution using biotinylated peptide representing the extracellular domain of the surface cancer associated protein AND competitive elution.

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Selection of Antibody Fragments scFv and VHH from phage display libraries for CAR construction

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