ProMab's custom rabbit monoclonal antibody development platform is parsed in four phases. As with every antibody generated, the aim of a ProMab Biotechnologies custom monoclonal antibody project is not only to generate an antibody that recognizes the immunizing antigen, but to raise an antibody that works for your specific application. ProMab's highly technical staff, with decades of experience spanning immunology, microbiology, biochemistry, cell and molecular biology will be there to guide you through the vital process of designing an appropriate antigen, and the subsequent, and equally important step of designing a strategy required for the generation of a truly successful, and validated antibody.
We provide detailed technical assistance in antigen design. In addition, we can also generate the antigen for you if you so prefer. Whether that is a peptide or recombinant protein (from one of three expression systems, including E. coli, baculovirus, or mammalian), we can make it for you.
Briefly, in phase I, one pair of New Zealand rabbits are immunized under our standard protocol (as shown below). Titers are evaluated after the third immunization. Once an acceptable titer is reached, we collect PBMCs from one or both rabbits for B cell sorting. We will keep the rabbits alive during the entire project.
In addition, you may choose to start with more than one pair of rabbits, or you may have an immunization protocol that you'd like us to use. We will accommodate most of your requests - just let us know!
Once appropriate ELISA positive titers are recorded toward the immunizing antigen, B cells are sorted by flow Cytometry using a dual-screening protocol that will ensure only rabbit lgG and antigen-specific B-cells are isolated (rabbit IgG antibodies do not show isotypes as found in mice and rats). Phase II then sees the isolated B cells used to generate a surface antibody display library. The library is then in Phase III through multiple rounds of cell panning to isolate cells bearing antigen-specific antibodies.
Isolated cells are then moved into phase IV, which sees the generation of a secreted recombinant antibody library, producing full, and complete rabbit monoclonal IgG antibodies. Following multiple-rounds of screening, candidate recombinant antibodies are expressed,purified,and validated for use in the applications of your choice.
ProMab's proprietary procedure will allow for identical immunization schedules to RabMab's generated using a standard fusion technique, but will then allow for an increased yield of antigen-specific B-cell candidates. Isolation of B-cells displaying antigen-specific antibodies will be followed by the production of a recombinant antibody library, allowing for not just the isolation of complete RabMab's, but also sequence information for both the Heavy and Light chains.
In addition to the accelerated delivery of important IP-related information, and therefore negating the requirement for additional hybridoma sequencing, as the production of your rabbit antibody(s), will be completely stable as expression plasmids, you will also be in the position of being able to generate an unlimited quantity lot-to-lot functionally stable antibody.