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Please join us for us for a Research Seminar Series talk by Aster Escalante, KGI Lecturer, Bioprocessing: Improving Expression and Downstream Purification of Insulin Molecules in Pichia pastoris by Alternative Promoters and Aqueous-Based Process Methodologies
In the next decade and beyond, global demand for insulin is expected to rise significantly requiring additional manufacturing capacity. The next generation of insulin manufacturing plants will likely be based on novel and robust platforms that are flexible, safe, simple to implement in a manufacturing setting and capable of step improvements in productivity and cost compared to current manufacturing techniques. Towards this end, Pichia pastoris expression systems have been evaluated for insulin production due to the organism’s capacity to secrete a variety of heterologous proteins and its ability to achieve high cell density cultures. Under the well-characterized, tightly regulated AOX1 promoter, inducible via methanol, yields of 1.5 to 3 g/L of purified insulin have been reported. However, methanol is a very volatile substance requiring specialized facilities, which can hamper large-scale production. Downstream processing of insulin precursors also requires use of organic solvents which can also burden manufacturing.
I will be discussing the development of an insulin process using a constitutive vs inducible promoter for protein expression and a simplified downstream purification process involving protein precipitation.