Abstract Title
Characterizing the Electrostatic Interactions Between Maltose Binding Protein and an Inflammatory Cytokine for Purification
Additional Funding Sources
This work presented here was supported in part by the National Center For Advancing Translational Sciences of the National Institutes of Health under Award Number UL1 TR002319. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Abstract
About 491,000 people are diagnosed with breast and prostate cancer every year12. Most cancer is treatable in its early stages; however, once the cancer cells metastasize (or spread to a secondary location), treatment options become limited. Small proteins called inflammatory cytokines play a role in regular inflammatory processes, but they can also signal cancer cells to metastasize. Collaborative research efforts are working to synthesize a small molecule inhibitor (SMI) capable of binding to an inflammatory cytokine (IC) to inhibit the signal responsible for metastasis. The inflammatory cytokine must be purified from a recombinant fusion protein to characterize the SMI binding site. Previous purification efforts have failed to produce high quantities of pure inflammatory cytokine due to the attraction between the inflammatory cytokine and maltose-binding protein (MBP). Modeling data supports that maltose-binding protein and the inflammatory cytokine exhibit electrostatic and hydrophobic interactions, causing difficulties separating the two molecules. The following workflow explores the effects of different chromatography methods to separate the inflammatory cytokine from maltose-binding protein.
Characterizing the Electrostatic Interactions Between Maltose Binding Protein and an Inflammatory Cytokine for Purification
About 491,000 people are diagnosed with breast and prostate cancer every year12. Most cancer is treatable in its early stages; however, once the cancer cells metastasize (or spread to a secondary location), treatment options become limited. Small proteins called inflammatory cytokines play a role in regular inflammatory processes, but they can also signal cancer cells to metastasize. Collaborative research efforts are working to synthesize a small molecule inhibitor (SMI) capable of binding to an inflammatory cytokine (IC) to inhibit the signal responsible for metastasis. The inflammatory cytokine must be purified from a recombinant fusion protein to characterize the SMI binding site. Previous purification efforts have failed to produce high quantities of pure inflammatory cytokine due to the attraction between the inflammatory cytokine and maltose-binding protein (MBP). Modeling data supports that maltose-binding protein and the inflammatory cytokine exhibit electrostatic and hydrophobic interactions, causing difficulties separating the two molecules. The following workflow explores the effects of different chromatography methods to separate the inflammatory cytokine from maltose-binding protein.