To improve our project we will compare our antibody scFv purification system with industrially used purification systems. OsmY has shown to be secreted in higher amounts than the industrially used pelB tag (Qian et al., 2008). In hopes that our purification system allows more scFv production we will compare the amount of scFv produced per liter of media using our construct to an industrially used construct.

One must also consider the limitation of our system. The scFv we used has already been secreted in E.coli and shown to function (Meyer, et al., 2012). It would be interesting for us to test a variety of antibodies to find which type of antibodies are easily excreted in E.coli with our construct. Perhaps a wider variety of antibodies can be secreted using our system than the common industrial system.

Our system isn’t constrained to antibodies; we could also use it for other disulfide bonded protein such as various cofactors, insulin, etc. Finally, in search of mimicking antibody’s agglutination properties it would be interesting to purify two antibody scFv linked together by a single peptide chain; enabling one construct to bind two antigens.

References:

Meyer, T. et al. Identification of immunogenic proteins and generation of antibodies against Salmonella Typhimurium using phage display. BMC Biotechnology 12, 29 (2012).

Qian, Z.-G. G., Xia, X., Choi, J. H. & my Lee, S. Y. Proteome-based identification of fusion partner for high-level extracellular production of recombinant proteins in Escherichia coli. Biotechnology and bioengineering 101, 587–601 (2008).