Walid Qoronfleh
Vice President, Business Development,
NextGen Sciences, Inc.
Abstract:
Over-expression of heterologous proteins in Esherichia coli is
commonly hindered by the formation of inclusion bodies.
Nevertheless, refolding of proteins in vitro has become an essential
requirement in the development of structural genomics )proteomics) and
as a means of recovering functional proteins from inclusion bodies.
Many distinct methods for protein refolding are now in use.
However, regardless of method used, developing a reliable protein
refolding protocol still requires significant optimization through trial
and error. Many proteins fall into the category of "Challenging"
or "Difficult to Express" and are problematic to refold using
traditional chaotrope-based refolding techniques. This review
discusses new methods for improving protein refolding, such as
implementing high hydrostatic pressure, using small molecule additives
to enhance traditional protein refolding strategies, as well as
developing practical methods for performing refolding studies to
maximize their reliability and utility. The strategies examined
here focus on high-throughput, automated refolding screens, which can be
applied to structural genomic projects.
