Novel eukaryotic cells and methods for recombinantly expressing a product of interest

Abstract

The disclosure pertains to novel eukaryotic cell suitable for recombinant production of a product of interest, wherein the effect of the expression product of an endogenous gene C12orf35 is impaired in said cell, thereby increasing their productivity when recombinantly expressing a polypeptide of interest. Furthermore, the present disclosure provides associated technologies wherein such host cells are used in recombinant production technologies.

Description

RELATED APPLICATIONS[0001]This application is a continuation of U.S. application Ser. No. 15 / 104,580, filed Jun. 15, 2016, which is a 35 U.S.C. 371 national stage filing of International Application No. PCT / IB2014 / 067073, filed Dec. 18, 2014, which claims priority to U.S. Application No. 61 / 919,313, filed Dec. 20, 2013. The entire contents of the aforementioned applications is incorporated herein by reference.SEQUENCE LISTING[0002]The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Nov. 4, 2021, is named NV1-098USCN_Sequence_Listing.txt and is bytes in size.FIELD OF THE DISCLOSURE[0003]The present disclosure concerns the field of recombinant expression technologies. It inter alia provides altered eukaryotic cells which are capable of enhanced production of a product of interest as well as their use in recombinant expression methods. Furthermore...

AI Technical Summary

Benefits of technology

The present patent is about a new discovery that a gene called C12orf35 affects the production of a recombinant protein in a cell. When this gene is impaired, the production of the recombinant protein is significantly increased. This finding is important because it helps to identify a key gene that can improve the efficiency of recombinant production. By reducing or eliminating the expression of C12orf35 in a cell, the yield of the recombinant protein is increased. This patent thus contributes to the field of genetic engineering.

Problems solved by technology

The generation of recombinant cell clones for production of therapeutic proteins usually comprises excessive screening of individual clones to detect and isolate high expressing clones.

However, even if high expressing clones are identified in the course of the screening process, these initially high expressing clones often lose their advantageous expression characteristics and the expression yield decreases over time.

This gradual loss of recombinant protein expression in cell clones during prolonged subculture is a common issue with many cell lines such as CHO cell lines and is referred to as instability.

This instability seriously affects the industrial production process of recombinantly produced polypeptides.

Therefore, the generation of recombinant cell clones for production of therapeutic proteins and other recombinant polypeptides that are produced on large scale usually comprises excessive and time-consuming screening of individual clones in order to identify the high expressing cell clones that also show the expression stability necessary for large scale production.

This practice prolongs the development of biotechnological such as biopharmaceutical production processes.

Even when using highly stringent selection systems that favour survival of high expressing cells under the used selection conditions, finding a suitable production clone within the surviving population which combines a high expression rate with good growth and stability characteristics is difficult.

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