Antibody development platform
A broad suite of antibody identification, optimization, engineering, characterization, and expression platforms.
The isolation of a lead antibody is an early and critical stage of CAR-T development. At CARsgen, we create antibodies using two well-established core technologies:
Hybridoma production basically relies on the fusion of immunized lymphocytes from an experimental animal with immortal myeloma cells. The resulting cell hybrid contains the genetic material of both parents. From the tumor cell the hybrid acquires the capacity for indefinite growth while the B lymphocyte confers the ability to synthesize a specific antibody. Hybridomas and the antibodies they produce can be selected and characterized for any given property.
This technology has been invaluable in generating several therapeutic molecules. CARsgen improves the pace and efficiency of this platform by developing proprietary immunization, fusion, and screening strategies.
In this technique, a diverse group of antibody genes prepared from a patient or generated in vitro are inserted into a phage coat protein gene causing the phage to “display” the antibodies on its outside while containing the gene for the antibodies on its inside. Antibodies displayed this way retain their ability to interact normally with other molecules, and our controlled selection processes of large libraries can identify hundreds of different potential CAR-T candidates to each target protein.
“Antibody optimization is a key process used to modify antibodies obtained from a primary antibody source, such as a murine hybridoma or human phage displayed antibody library, into molecules that possess properties desired in safety, biophysical, and functional properties, or potency.“
— Dr. Zonghai Li, President&CEO
For reducing the potential risk of immunogenicity associated with animal-derived antibodies, antibody humanization is an essential technology. Humanization consists of grafting the antibody’s complementary determining regions (CDRs) onto carefully selected full-length human frameworks while the critical properties of the antibody that should be sufficiently qualified, including immunogenicity, binding affinity, physiochemical stability.
At CARsgen, we can often achieve optimization of the lead antibody’s specific activity by using an affinity maturation process. Typically, in vitro affinity maturation consists of two well-defined components:
A strategy to generate libraries of antibody variants and a selection or screening strategy to enable isolation of variants of interest. The strategies to generate libraries of antibody variants include random and rational mutagenesis. Rational strategies generate diversity in predefined segments of the antibody V region, whereas random strategies introduce point mutations, insertion/deletions or homologous fragments containing more than one amino acid replacement all along the V region.
Structural biology efforts can help to provide atomic level insights into structure-function relationships and the antibody-antigen interaction, inform future strategies for antibody optimization. At CARsgen, epitope mapping, computational modeling are often used as major approaches.