Both breast and prostate tumors can be surgically removed when in Stages I, II, and III. Once the tumor undergoes metastasis (stage IV), survival rates drop; and the surgical removal of the primary tumor will no longer be sufficient for treatment. An inflammatory cytokine (IC), a protein that acts as a signal molecule in inflammation, has been identified as a primary signal in metastasis. Therefore, targeting the inflammatory cytokine with a small molecule inhibitor (SMI) will prevent metastasis from occurring. Structure-based drug design requires an accurate measurement of how well the SMI is able to bind to the inflammatory cytokine. This binding requires the expression and purification of the inflammatory protein with high purity and accuracy. The construct designed for the purification consists of the cytokine protein attached to a maltose-binding protein (MBP) and six histidines. The MBP serves to increase solubility; however, it also is electrostatically attracted to the inflammatory cytokine. To ensure efficient separation of these two proteins, a six-histidine tag is attached to the MBP. Ongoing experiments aim to design a protocol to produce pure, bioactive protein.
Rushing, Brittany; Delamontanya, Katelyn; Engmann, Terrell; Suman, Hanna; Grantham, Bri; Baggs, Eric; and Warner, Lisa, "Expression and Purification of Inflammatory Cytokine Protein" (2021). 2021 Undergraduate Research Showcase. 48.