Cell-free protein synthesis (CFPS) is an alternative method to cellular expression for recombinant protein production.
Its lack of cellular boundaries enables direct modification of the CFPS reaction to allow optimised and rapid production of functional and labelled proteins.
Results indicate the p H and Temperature dependent degradation of Engineered Proteins by collagenase require investigation into other degradable sequences as substitutes for current CLS to better improve lifetime in inflammatory conditions.
Also, future research into kinetic profiles of the degradation of PCLP and CCLP by MMPs are necessary for optimizing the lifetime of the hydrogel for in-situ drug delivery.
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Theses and dissertations represent a wealth of scholarly and artistic content created by masters and doctoral students in the degree-seeking process.Full-length CXCR4 and CD4 were produced in proteoliposomes or additionally, for CXCR4, as detergent solubilised protein.However, the expression and purity levels of solubilised CXCR4 and CD4 remained too low to permit advanced structural work.Pathologically, CXCR4/SDF1-α are involved in cancer progression, and CD4 is the primary receptor for HIV GP120 (glycoprotein 120) during viral infection.For the purposes of integrated structural biology, a primary objective was to optimise CFPS to generate sufficient amounts of protein.Pleiotropic effects from oral administration of anti-inflammatory drugs limit their effectiveness.The ultimate goal of this project is to develop a novel self-assembling protein-based hydrogel for in situ delivery of NSAIDs at the site of chronic inflammation.This important aspect of this responsive material requires susceptibility to relevant proteases, and is the specific subject of the present study.Exposure of CCLP to collagenase, and MMP-1, indicates that each protomer is susceptible to cleavage in the CLS sequences.High-cell density cultures (HCDC) in fermenters were then established for lysate production.Subsequently, HCDC was used for the production of deuterated lysates for SANS (small-angle neutron scattering) studies.