Cell line for producing adenovirus and method of preparing the same
Inventors
Assignees
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Publication Number
US-11299713-B2
Publication Date
2022-04-12
Expiration Date
Abstract
The present invention relates to a cell line for producing an adenovirus having a limited autoreplication capability, and a method of preparing the same, and more particularly, to a cell line for producing an adenovirus having no autoreplication capability by expressing at least one selected from an adenoviral E1 protein and an E1A or E1B protein, and a method of preparing the same. Also, the present invention relates to the use of the cell line expressing at least one selected from the adenoviral E1 protein and the E1A or E1B protein.
Core Innovation
The invention provides a cell line for producing an adenovirus that comprises an E1 gene of 2962 bp including both an E1A gene and an E1B gene, and further comprises an E1A gene of 998 bp separate from the E1A gene of the 2962 bp E1 gene, an E1B gene of 1808 bp separate from the E1B gene of the 2962 bp E1 gene, or both. The cell line does not generate replication competent adenovirus (RCA).
The invention also provides a method of preparing a cell line for producing an adenovirus. The method prepares a first retrovirus vector comprising an E1A gene of 998 bp, a second retrovirus vector comprising an E1B gene of 1808 bp, and a third retrovirus vector comprising an E1 gene of 2962 bp including both the E1A gene and the E1B gene, and then uses these retrovirus vectors to obtain a cell line that comprises the specified E1 gene architecture and does not generate replication competent adenovirus (RCA).
The invention further provides downstream compositions that use an adenovirus obtained from the RCA-non-generating cell line. A packaging composition for adeno-associated virus (AAV) production includes the adenovirus and a cell line comprising a nucleic acid sequence of a rep gene of the AAV and a nucleic acid sequence of a cap gene of the AAV, and includes an AAV vector. A composition for gene therapy comprises the adenovirus obtained from the RCA-non-generating cell line and is defined by the same E1 gene architecture constraints.
Claims Coverage
The provided claim set includes three independent claims, covering a cell line, a method, and compositions. Across these claims, the inventive scope is centered on an adenovirus-producing cell line with a specific E1 gene arrangement and the limitation that the cell line does not generate replication competent adenovirus (RCA).
RCA-non-generating adenovirus-producing cell line with separated E1A/E1B components
A cell line for producing an adenovirus comprising an E1 gene of 2962 bp including both an E1A gene and an E1B gene, and an E1A gene of 998 bp separate from the E1A gene of the 2962 bp E1 gene, an E1B gene of 1808 bp separate from the E1B gene of the 2962 bp E1 gene, or both, wherein the cell line does not generate replication competent adenovirus (RCA).
Retrovirus-vector preparation of the RCA-non-generating adenovirus-producing cell line
A method of preparing a cell line for producing an adenovirus by preparing a first retrovirus vector comprising an E1A gene of 998 bp, a second retrovirus vector comprising an E1B gene of 1808 bp, and a third retrovirus vector comprising an E1 gene of 2962 bp including both the E1A gene and the E1B gene, and using these retrovirus vectors to obtain the cell line that comprises the specified E1 gene architecture and does not generate replication competent adenovirus (RCA).
AAV packaging composition using adenovirus from the RCA-non-generating cell line with defined E1 architecture
A packaging composition for adeno-associated virus (AAV) production comprising an adenovirus obtained from a cell line that does not generate replication competent adenovirus (RCA) and comprises an E1 gene of 2962 bp including both an E1A gene and an E1B gene and an E1A gene of 998 bp separate from the E1A gene of the 2962 bp E1 gene and an E1B gene of 1808 bp separate from the E1B gene of the 2962 bp E1 gene, or both; a cell line comprising a nucleic acid sequence of a rep gene of the AAV and a nucleic acid sequence of a cap gene of the AAV; and an AAV vector.
Gene therapy composition using adenovirus from the RCA-non-generating cell line with defined E1 architecture
A composition for gene therapy comprising an adenovirus obtained from a cell line that does not generate replication competent adenovirus (RCA) and comprises an E1 gene of 2962 bp including both an E1A gene and an E1B gene and an E1A gene of 998 bp separate from the E1A gene of the 2962 bp E1 gene and an E1B gene of 1808 bp separate from the E1B gene of the 2962 bp E1 gene, or both.
The claims focus on engineering an adenovirus-producing cell line with a defined E1 gene architecture and the limitation that the cell line does not generate replication competent adenovirus (RCA), and on using the resulting adenovirus in AAV packaging and gene therapy compositions.
Stated Advantages
The cell line does not generate replication competent adenovirus (RCA).
Documented Applications
Packaging composition for adeno-associated virus (AAV) production.
Composition for gene therapy.
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