To diversify the functions and properties of hybridoma produced antibodies, genome engineering using CRISPR/HDR is used.
Bioengineers, life scientists and biochemists use hybridoma to create large quantities of identical antibodies and to develop new diagnostics and therapeutics. Recent preclinical studies and clinical trials on the technology have highlighted the importance of isotypes in antibody therapeutics. A research team from the Netherlands has developed a CRISPR and HDR platform that allows them to quickly engineer immunoglobins domains, and create recombinant chimeras or mutants with designer antibodies in a desired format, species, or isotype. Johan M. S. van der Schoot, along with colleagues from the departments of immunology (immunohematology), translational immunology, medical oncology and proteomics (proteomics), used this platform to create recombinant mutants, chimeras, and hybridomas. The antigen-specificity of the stable antibodies was retained. The team believes that the versatile platform will enable mass-scale antibody manufacturing for the scientific community in order to advance preclinical antibody studies. Science Advances has published the work.
The monoclonal antibody (mAb), which was developed to treat diseases once considered incurable, has revolutionized medical science. Hybridomas have been used for decades to discover, screen and produce mAbs. They are immortal cell lines capable of producing large quantities of mAbs that can be used for new antibody-based treatments. In the last decade, scientists had created, validated, and assisted a large number hybridomas for preclinical studies, where the mAb isotypes and format were crucial to understanding their performance in preclinical model. mAbs that are genetically engineered are usually produced using recombinant technologies, in which the variable domains must be sequenced, copied into plasmids, and expressed on transient systems. These processes are difficult, time-consuming and expensive. They lead to the outsourcing of work to contract research companies. This hinders academic research and early-stage antibody research.
The constant antibody domains that form the fragment crystallizable- (Fc), domain, are crucial to the therapeutic efficacy mAbs because they interact with specific FcRs. Prior research had highlighted Fc’s central role in antibody-based therapies to highlight this role. Since its advent, CRISPR and associated protein Cas-9 (CRISPR-Cas9)-targeted genome editing technology has opened multitudes of exciting opportunities for gene therapy, immunotherapy and bioengineering. Researchers used CRISPR Cas9 to modify mAb expression within hybridomas, create a hybridoma-platform and engineer hybridomas for antibody modification. It is still necessary to genetically engineer a platform that allows for effective and versatile Fc replacement from foreign species in hybridomas.
Source:
https://phys.org/news/2019-09-genome-crisprhdr-diversify-functions-hybridoma-produced.html
