Expression of heterologous proteins

Abstract

An expression system which provides heterologous proteins expressed by a non-native host organism but which have native-protein-like biological activity and / or structure. Disclosed are vectors, expression hosts and methods for expressing the heterologous proteins. The expression system involves co-expression of protein factor(s) which is / are capable of catalyzing disulphide bond formation and desired heterologous protein(s). The expression system is presented using yeast cells as the preferred host, protein disulphide isomerase (PDI) and thioredoxin (TRX) as the preferred examples of the protein factors and HCV-E2715 envelope glycoprotein and human FIGF as the preferred examples of the heterologous proteins.

Description

[0001] The present invention relates to an expression system which provides heterologous proteins expressed by a non-native host organism but which have native-protein-like biological activity and / or structure.BACKGROUND TO THE INVENTION[0002] Advances during the past decade in molecular biology and genetic engineering have made it possible to produce large amounts of protein products using heterologous expression systems.[0003] The use of heterologous hosts for production of, for example, therapeutic proteins, can lead, however, to differences in the biological and / or structural properties of the recombinant product. Amongst the biochemical modifications that commonly occur to proteins during or following their synthesis in the cell, the formation of disulphide bonds is of relevance since this modification is coupled to the correct folding or assembly of disulphide-bonded proteins (reviewed by J. C. A. Bardwell and J. Beckwith, Cell, 74: 769-771, 1993; R. B. Freedman, in Protein Fo...

AI Technical Summary

Problems solved by technology

The use of heterologous hosts for production of, for example, therapeutic proteins, can lead, however, to differences in the biological and / or structural properties of the recombinant product.

In the numerous organisms, for example, bacteria, yeast, mammalian cells and insect cells, which have been genetically manipulated to (over)express heterologous proteins, the problem encountered with most expression systems is the inability to express proteins which are biologically active.

It now appears that due to the lack of or inefficient amount of the enzymes necessary for correct folding or assembly of heterologous proteins in non-native expression hosts, such expressed heterologous proteins are often not biologically active and / or have an incorrect protein structure.

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