Research target:
New hotspots in the field of antibody drug targets: tumor immunotherapy, programmed death receptor 1 (PD-1)
Programmed death receptor 1 (PD-1) is an important immunosuppressive receptor molecule expressed in activated B cells, T cells and bone marrow cells, with PD-1 as a target for immune regulation in anti-tumor and anti-tumor Infection, anti-autoimmune diseases and survival of organ transplants are all important.
Thesis:
As one of the immunological checkpoint proteins, PD-1 plays an important role in down-regulating T cell activation. By binding to the ligands PD-L1 and PD-L2 expressed on the surface of tumor cells, it inhibits T cell proliferation by binding inhibitory signals. Thus, the tumor cells escape the surveillance and attack of the human immune system. Therefore, PD-1 is an important target in cancer immunotherapy. PD-1 immunotherapy is to design specific protein antibodies against PD-1 or PD-L1, prevent the recognition process of PD-1 and PD-L1, partially restore T cell function, so that T cells can kill tumor cells and relieve tumors. Cells evade the immune system. PD-1 antibody is currently a type of cancer therapy that has received much attention. Although therapeutic antibodies are developed for humans, in the preclinical phase, the efficacy and safety of the molecules need to be evaluated in animal models. Among them, the most direct and most cost-effective method is to test antibodies that cross-react with both human and murine targets in a mouse model.
Given the importance and widespread use of PD-1 antibodies in the treatment of various cancers, this study hopes to develop such antibodies to accelerate preclinical studies of PD-1 antibodies. PD-1 antibodies currently approved for clinical or ongoing development are only targeted to human hPD-1 and do not appear to cross-react with murine mPD-1. Therefore, this study developed and reported for the first time a functional antibody with a human/mouse PD-1 cross-reactivity and compared the epitope of this antibody to an antibody epitope that only binds to hPD-1.
In this study, rats were first immunized with hPD-1 and mPD-1, and a large number of rat hybridoma clones were screened to find an antibody R9 that targets both hPD-1 and mPD-1 and only targets hPD. -1 antibody R11. To quickly identify the epitopes of the two antibodies, the study used alanine scanning and was based on ELISA and surface plasmon resonance techniques. At the same time, combined with the molecular simulation method, the R11 and R9 antibody structural models were constructed by using the MODELER program in the Model Antibody tool in Discovery Studio, and based on this, the ZDOCK/RDOCK protein-protein docking/optimization program in Discovery Studio was used. Two antibodies were ligated to hPD-1, predicting different binding patterns of the two antibodies, and studying their binding epitopes to explain the cross-reactivity mechanism of human/mouse cross-reactive antibodies.
Figure. Prediction of binding patterns of antibodies R11 (A), R9 (B) and hPD-1.
The Discovery Studio module used in the antibody simulation section of this article:
· DS MODELER: An industry-standard homology modeling program developed by professor Sali in 1993. The program has been cited more than 9,000 times in academic papers, and academic results are available in various magazines. As long as a protein with an unknown structural sequence and another known three-dimensional structure with some homology is given, the protein structure can be predicted automatically and quickly. In addition to the universal protein homology modeling module, there are specialized antibody modeling modules that enable prediction of antibody Fab regions, Fv regions, full-length antibodies, and bispecific antibody structures.
· DS Protein Docking: A fast and accurate protein-protein (antibody-antigen) complex structure prediction tool. Protein Docking uses the classic and reliable ZDOCK algorithm to predict the structure of protein-protein complexes, including ZDOCK, ZRANK and RDOCK. ZDOCK is used for protein-protein rigid docking, ZRANK is used for energy-based re-scoring of ZDOCK-derived structures, and RDOCK is optimized for ZDOCK based on CHARMm energy minimization.
Other modules in Discovery Studio that can be applied to the design of new antibody products can be found at: http://uee.me/dG6u "Discovery Studio Application Solutions in Antibody R&D"
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