Sulfitolysis efficiency in downstream processing of recombinant human insulin production and estimating the secondary structure by circular dichroism (CD)

During the production of recombinant human insulin, before final refolding, the S-S bonds need to be opened in a process of oxidative breakdown called sulfitolysis. In this process, sulfur atoms are converted into –SO3-. Sulfitolysis is a crucial step in the whole downstream process as it can produce the higher amount of properly folded fusion protein feeding the later digestion process; therefore make it vulnerable to higher scrutiny and optimizations. Circular dichroism is a top of the line instrumentation technology to elucidate the secondary structure of the proteins and peptides. Using this technique would help to not only compare the structural manipulations in a step-wise process, but also to quantify the elements of the secondary structure in each step. This method proved to be an effective downstream method optimizing assistant while having the capability of being routinely used as a complementary QC instrument, which can identify the nature of peptide folding in each step.

کلیدواژه ها:

fusion protein ، circular dichroism ، human insulin ، folding ، downstream processing