Systems and methods for expressing proteins

Abstract

The present disclosure relates to method and compositions for generating proteins. In particular, the present disclosure relates to electroporation mediated gene delivery in the generation of recombinant proteins (e.g., drug metabolizing enzyme and transporter vesicles) in mammalian cells.

Description

[0001]This application claims the benefit of priority under 35 U.S.C. § 119 of U.S. Provisional Application Ser. No. 62 / 259,788 filed on Nov. 25, 2015, the content of which is relied upon and incorporated herein by reference in its entirety.FIELD OF THE DISCLOSURE[0002]The present disclosure relates to method and compositions for generating proteins. In particular, the present disclosure relates to electroporation mediated gene delivery in the generation of recombinant proteins (e.g., drug metabolizing enzyme and drug transporter vesicles, microsomes or cell fractions) in mammalian cells.BACKGROUND OF THE DISCLOSURE[0003]Expression of recombinant proteins is important in many aspects of the research, industrial, and pharmaceutical biotechnology industries. For example, the expression of drug metabolizing enzymes and transporter proteins is often critical in drug discovery and development. For many of these proteins, expression in mammalian cells is preferred over expression in proka...

AI Technical Summary

Benefits of technology

The present patent is about a method for generating proteins in mammalian cells using electroporation-mediated gene delivery. This method can be used to generate membrane-bound proteins, such as drug metabolizing enzymes and drug transporters, in vesicles or microsomes. The method involves contacting a mammalian cell with a nucleic acid encoding the protein of interest, and then isolating the membrane-bound protein from the cell. The isolated membrane-bound protein can be used for various applications such as drug screening and inhibitor development. The patent also provides isolated vesicles or microsomes generated by the method.

Problems solved by technology

Such problems include cell toxicity, inability to deliver larger genes or genetic constructs, significant batch-to-batch inconsisteny in expression levels, unstable expression, and improper localization, post-translational modification, and / or folding of expressed proteins.

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