Harnessing IMAC To Enable An Immobilized Enzyme Cascade For The Synthesis Of Islatravir

Advances in enzyme engineering have dramatically increased the use of novel enzyme classes and their applications to pharmaceutical synthesis. In many instances, multiple enzymes are simultaneously employed for co-factor recycling or to perform sequential synthetic steps. The high enzyme loads for these processes introduce many challenges related to protein stability and protein-product isolation, which are traditionally mitigated by process-intensive operations with significant yield loss. Novel strategies are needed to increase enzyme loading, stability, remove residual host cell proteins with off-target activity, and to facilitate product-protein separation without additional chemical isolations or purifications.

Immobilizing an enzyme to a porous resin solves many of these issues by creating a solid-supported catalyst with extremely high enzyme loads that can be prepared in advance, easily charged into a reactor, and rapidly isolated from the product by filtration. We have developed a scalable enzyme-immobilization platform technology based on IMAC to selectively co-immobilize enzyme cascades onto a single resin directly from crude cell lysate and reject host-cell proteins. In this talk, Jacob H. Forstater, Ph.D., Associate Principal Scientist, Merck Process Research & Development at Merck Research Laboratories will discuss how we utilized IMAC to enable a  streamlined through-process for the biocatalytic synthesis of islatravir, a promising nucleoside analog drug for the treatment of HIV. We discuss process design principles for the development and scale-up of robust multi-enzyme biocatalytic processes and their integration into traditional small-molecule manufacturing operations.  

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