A team of researchers has successfully incorporated hybridoma technology with CRISPR/HDR to produce large numbers of identical antibodies. This could have huge implications for future antibody-based therapeutics and diagnostic techniques.
This highly versatile new technique, the team believes, should facilitate mass-scale antibody engineering for the scientific community. It will empower preclinical antibody research.
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In a new paper released in the August edition of the journal Science Advances, a team of scientists has made a breakthrough in hybridoma-produced antibodies using CRISPR/HDR.
Johan M. S. van der Schootet al., believe this novel antibody production process could have major implications for treating some debilitating diseases in the future.
"Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted," states the research team in the research abstract.
By integrating CRISPR (Clustered Regularly Interspaced Short Palindrome Repeats) and HDR (homology-directed repair) techniques, they have been able to create a new method that allows for the rapid engineering of constant immunoglobulin domains to obtain recombinant hybridomas. These hybridomas secrete antibodies in the desired format, species, or isotope.
Hybridomas are hybrid cells that have been artificially created to produce large amounts of antibodies for diagnostic and therapeutic use.
"Hybridomas are produced by injecting a specific antigen into a mouse, collecting an antibody-producing cell from the mouse's spleen, and fusing it with a tumor cell called a myeloma cell," according to medicinenet.com.
What did the research team do?
The team used CRISPR/HDR to form recombinant hybridomas, chimeras, and mutants. These hybridomas were able to pump out monoclonal antibodies (mAb) of the type that have previously revolutionized the treatment of once incurable diseases, like some forms of cancer.
"Using this platform, we obtained recombinant hybridomas secreting Fab′ fragments, isotype-switched chimeric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemoenzymatic modification," states the published article.
The use of hybridomas to create mAbs is nothing new and has been used since the mid-1970s. However, these older techniques are time-consuming, challenging, and expensive. Parts, or all of the process, needs to be outsourced to contract research companies, which hampers the process of academic early-stage antibody development and preclinical research.
Not only is the new process quicker and relatively cheaper, but it also has a near 100% success rate.
This will prove critically important for preclinical studies where traditional-hybridoma products are often used in vivo. This new method should, the authors believe, empower clinical antibody research for therapeutic antibody development.
"We believe that this versatile platform facilitates antibody engineering for the entire scientific community, empowering preclinical antibody research," declares van der Schoot, et al.
For example, the site-specific functionalization of engineered antibody products will have wide-ranging applications in the fields of biomedical engineering, chemical biology, drug development, and nanomedicine. Not to mention having other potential applications for the entire scientific community.
The original paper was published in the journal, Science Advances.