The Biotechnology Core applies the practices of Molecular Biology and Protein Engineering for the expression and production of recombinant proteins and antibodies in both mammalian and E. coli expression systems. The Core is fully equipped for purification of the recombinant products via affinity and classical chromatography techniques, with rigid quality assurance analytics to verify protein activity and purity as well as freedom from endotoxin contamination.
An important function of the Core is the development of high-quality monoclonal antibodies against antigens of specific scientific interest to BIIR programs. Various applications for such antibodies are: 1) antigen detection via ELISA, flow cytometry, and microscopy; 2) biological activation or neutralization of critical pathway processes; and 3) development of antibody pairs that can be used as reagents for detecting and measuring cytokines. One use of such antibody-detection reagents is the high sensitivity and simultaneous measurement of analytes (such as cytokines/chemokines present in bodily and tissue culture fluids) using multiplexed bead-based Luminex technology. The Core hosts the Institute’s multiplexed bead-based cytokine assay service and participates in the EQAPOL (External Quality Assurance Program Oversight Laboratory) funded by NIH, NIAID, and DAIDS, for proficiency testing and compliance. The Core has developed antibody pairs representing unique cytokines (not commercially available) as well as commonly used cytokines for ‘cost saving’ immune-monitoring programs. The Core has also developed and provides multiplexed bead-based antigen-specific antibody assays for screening antibody titers from in-vivo mouse and NHP models, as well as for human clinical trials.
The core purifies monoclonal antibodies from the hybridomas generated in these development programs. The core provides 1) direct labeling of pure antibodies; 2) molecular cloning, sequence confirmation, and CDR identification of the functional antibody heavy and light chain variable domains from selected hybridoma clones; and 3) molecular construct design, cloning, and production associated with recombinant antibody generation.
In addition, the Core supports the DC-targeting vaccine programs through development of stable CHO-S cell lines expressing diverse anti-DC receptor-antigen fusion proteins. These prototype vaccines are provided for use in preliminary research investigation as well as in-vivo mouse and non-human primate animal models both here at BIIR and through international collaborative research agreements.