Optimization Protocol for Extraction and Purification of Recombinant Protein Modeled with Taq Polymerase


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This experiment explores recombinant protein drug protocols and how to better optimize and purify large scale recombinant proteins expressed in bacterial cells. Using Escherichia coli bacteria, the gene Taq will be cloned into the bacterial plasmid, grown, induced, and incubated . Taq proteins will then be transcribed, lysed out of cells, and purified using an ion exchange chromatography column. By altering elements of this current process — varying the column flow rate and using different salt gradient concentrations for column purification — more concentrated yields were achieved with greater purity. It was found that lower flow rates of 100 mL/hour at normal salt gradient concentration of 25 to 700 mM produced the most optimal protein fractions. While Taq polymerase is not a recombinant protein drug, it can serve as a model to represent therapeutic protein drugs and purification optimizations that would work well for both. Utilizing these results, they can potentially provide the pharmaceutical industry novel ways to increase their production while lowering costs. Drug manufacturers would be able to produce these drugs at higher rates and more affordable prices, while saving on resources and time. Patients purchasing recombinant protein drugs would benefit from this as drugs become more widely accessible and reasonably priced. Optimization of protein purification can overall ease economic burdens, and initiate change from typical protocols.



Recombinant protein, Eschericia coli, Taq polymerase, Ion exchange column chromatography, Protein extraction, Protein purification, Optimizing protocol