Selecting specificities for a Bispecific T-cell Engaged Antibody
A recent patent filed by Janssen exploited the potent antitumor functions of gamma delta T cells through the development of bispecific T-cell engaged antibodies. The gamma delta (γδ) T cells mainly express heterodimers of TRGVγ9 and Vδ2 chains, and the majority, if not all, of these cells exhibit efficient cytotoxicity of target tumor cells. Janssen harnessed this ability using bispecific antibodies to construct a BiTE such that one arm binds to the TRGV9 structure and the other arm binds to a T cell Tumor-Associated Antigen (TAA2) expressed by the tumor cells. Thus, the bispecific antibody bridges the effector and target cells together, resulting in tumor cell killing.
Engineering a Bispecific T-cell Engaged Antibody with the help of de novo antibody protein sequencing
In order to engineer this bispecific, Janssen needed to modify its sequence, but without access to a DNA sequence, it was difficult to proceed. Since they did have a protein sample, they took advantage of Rapid Novor’s REmAb® de novo antibody protein sequencing platform. To design the bispecific, they worked from two antibody sequences which they obtained by sequencing two monoclonal antibodies through REmAb®.
Thanks to our full coverage and high accuracy, we delivered excellent results. The sequences for the two antibodies had a minimum of 30 times coverage over every single amino acid, and hundreds of overlaps, which is why we could confidently claim 100% accuracy. Janssen expressed and purified the sequences of both antibodies. They found that the anti-TCR Vγ9 antibody bound to human γδT cells, as expected, and showed high specificity for TCR Vγ9. They also recapitulated similar results for their anti-TAA2 clone.
Safeguarding discoveries with de novo antibody protein sequencing
Using the two aforementioned in-house validated sequences, they engineered the anti-TRGVγ9/anti-TAA2 bispecific human IgG1 antibody. Following standard in vitro functional screening tests such as T-cell re-directed killing of TAA2-expressing H929 cells, and were able to prove that the BiTE was capable of mediating T-cell cytotoxicity against tumor cells.
At Rapid Novor, we believe that de novo antibody protein sequencing can advance the discovery of therapeutics and diagnostics, but it can also be used to safeguard these discoveries through patents. As other companies have and continue to trust us in the development of patents and publications, we will continue to innovate and ensure that next generation protein sequencing is accessible to researchers to enable their discovery.