Location: Commodity Utilization ResearchTitle: Improving recombinant protein purification yield) Author
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 3/12/2010
Publication Date: 3/21/2010
Citation: Cao, H. 2010. Improving recombinant protein purification yield (abstract). BIT Life Sciences' PepCon-2010 Conference, March 21-23, Beijing, China. p. 207. Interpretive Summary:
Technical Abstract: Production of adequate amounts of recombinant proteins is essential for antibody production, biochemical activity study, and structural determination during the post-genomic era. It’s technologically challenging and a limiting factor for tung oil research because analytical reagents such as high quality antibodies and pure proteins are not available. Some of the limiting factors in protein production include low level expression, precipitation, and loss of activity during the purification processes. This presentation will summarize the results on recombinant protein production from our current research on tung oil biosynthetic enzymes and previous research on maize starch biosynthetic enzymes and mammalian proteins. We propose several approaches to improving recombinant protein purification yield, including 1) selecting high level expression systems, 2) optimizing expression conditions, 3) promoting protein solubility, 4) minimizing purification steps, and 5) maintaining protein structure and function. Recombinant proteins/enzymes to be discussed are soluble and membrane-bound proteins involved in signal transduction and metabolic pathways. These proteins are expressed as tagged and native proteins in E. coli, yeast, and mammalian cells. The recombinant proteins are further purified using affinity and conventional chromatographic procedures. The purified proteins are subsequently characterized and utilized for antibody production, biological activity assays, and post-translational modification site identification by immunological, enzymatic, protein-RNA interaction, and mass spectrometry methods.