Novel Conjugation Sites for Antigen Binding Reagents

"This project proposes to cost effectively prepare scFv or Fab antibody fragments that have been modified to site-specifically incorporate stable linkable moieties that will allow their conjugation to reporter molecules and surfaces without comprising thebiological function of the binding site of the antibody fragment. The ready availability of technologies to prepare these antibody fragments with stable linkable moieties will allow manufacture of instruments for the sensitive detection and quantificationof analytes present in both biological and non-biological samples. We propose to engineer the proprietary hydrazine/carbonyl bioconjugation couple developed by Solulink to accomplish this task. As monoclonal antibodies (MoAbs) are expensive to manufactureand typically involve complex mammalian cell culture systems the use of transgenic expression systems, especially plants, can reduce these costs dramatically but they are unproven and take many years of development to produce just one antibody.Additionally, the manufacturing cycle for both mammalian cell culture and transgenic systems is long and involved. On the other hand, bacterial expression systems for the manufacture of scFvs and Fab antibody fragments are both inexpensive and quick todevelop. A viable E. coli fermentation system can be designed, engineered and developed in just a few days. Versatile, robust technologies for the production of antibody fragments that can be r