Chemical component limited culture medium for HEK293 cell culture and replication and amplification of adenovirus and adeno-associated virus

Abstract

The invention discloses a chemical component limited culture medium for HEK293 cell culture and replication and amplification production of adenovirus and adeno-associated virus. The chemical component limited culture medium consists of amino acid, vitamins, trace elements, inorganic salt and fatty acid. The chemical components of the culture medium are limited, the culture medium is clear in chemical element components and free of serum and protein, plant hydrolysate is not added, small molecular compounds with known structures and functions are adopted for substitution, HEK293 cell culture metabolism and replication and amplification of adenoviruses and adeno-associated viruses are more facilitated, and the culture medium is safer and more effective.

Description

technical field [0001] The invention belongs to the field of adenovirus, adeno-associated virus and bioengineering, and particularly relates to a chemical-component-limited medium suitable for HEK293 suspension large-scale amplifying culture and adenovirus and adeno-associated virus replication and amplification production. Background technique [0002] Adenovirus and adeno-associated virus (AAV) are two different types of viral vectors commonly used for gene delivery. These two recombinant virus systems have obvious advantages in infecting the host, such as low immunogenicity and infecting dividing and non-dividing cells. The differences between the two are mainly in the packaging capacity and level, the onset and duration of gene replication and amplification, and the difference in immune response. Adenoviruses have a capacity of about 8.5 kilobases and can replicate high levels of proteins and transient genes. The packaging capacity of AAV is about 4.5 kilobases, the le...

AI Technical Summary

Problems solved by technology

[0007] 1. The formula contains proteins, protein hydrolyzates and other substances that are not chemically limited, which makes the final product less safe;

[0008] 2. The medium with non-chemically limited components has insufficient stability, and it is difficult to control the batch-to-batch difference during the large-scale production of the medium;

[0009] 3. The non-chemically restricted substances contained are sensitive to the environment, resulting in harsh conditions for the transportation and storage of culture medium products;

[0010] 4. Substances such as protein or protein hydrolyzate affect the separation and purification of the final product, and are difficult to remove in the downstream separation and purification process. The requirements for separation and purification are high, the cleaning of the purification column is difficult, the separation and purification time is prolonged, and the service life of the purification column is reduced;

[0011] 5. Full formula grinding cannot effectively guarantee product quality. Most products contain multiple components such as basal medium and additives, which cause inconvenience to product transportation and storage and customer liquid preparation;

[0012] 6. The medium product with limited chemical composition is single, and there is no corresponding development for the R&D and production of adenovirus and adeno-associated virus. After virus transfection, the activity and continuous amplification ability of cells decrease, resulting in insufficient product titer

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