The Rutgers antigen production group is responsible for designing, expressing, purifying, and shipping protein domain antigens to the Protein Capture Reagents Program antibody production centers at CDI/JHU and RAN. The initial set of human transcription factors includes targets nominated by the community, as well as annotated transcription factor proteins from various databases. Customized bioinformatics software is used to parse these multidomain proteins into constructs corresponding to a single well-folded domain, which are then synthesized, cloned, and expressed recombinantly in E. coli.
The transcription factor domains are purified by IMAC and GF, assayed for QA/QC, and buffer exchanged into the desired formulation. An N-terminal AviTag allows for site-specific biotinylation of the antigens if required. To date, the Rutgers antigen production group has delivered over 2000 protein samples representing nearly 1000 unique human transcription factors.