Cell line for producing adenovirus and method of preparing the same

A method of preparing a cell line for producing an adenovirus, the method comprising:

• (a) preparing i) a first retrovirus vector comprising an E1A gene of 998 bp or an expression product thereof, ii) a second retrovirus vector comprising an E1B gene of 1808 bp or an expression product thereof, and iii) a third retrovirus vector comprising an E1 gene of 2962 bp including both the E1A gene and the E1B gene or an expression product thereof;
• (b) transfecting the first, second, and third retrovirus vectors into cell lines for producing retroviruses to thereby obtain a first retrovirus, a second retrovirus, and a third retrovirus, respectively;
• (c) infecting a cell line for producing an adenovirus with the third retrovirus;
• and
• (d) further infecting the cell line for producing an adenovirus with the first retrovirus, the second retrovirus, or both, to thereby obtain the cell line that (1) comprises:
• i) an E1 gene of 2962 bp including both an E1A gene and an E1B gene;
• and
• ii) an E1A gene of 998 bp that is separate from the E1A gene of i) above, an E1B gene of 1808 bp that is separate from the E1B gene of i) above, or both,
• wherein the cell line is capable of producing the adenovirus, and does not generate replication competent adenovirus (RCA).

A packaging composition for adeno-associated virus (AAV) production comprising:

• an adenovirus obtained from a cell line for producing an adenovirus wherein the cell line does not generate replication competent adenovirus (RCA) and the cell line comprises:
• i) an E1 gene of 2962 bp including both an E1A gene and an E1B gene;
• and
• ii) an E1A gene of 998 bp that is separate from the E1A gene of i) above, an E1B gene of 1808 bp that is separate from the E1B gene of i) above, 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.

A composition for gene therapy comprising

• an adenovirus obtained from a cell line for producing an adenovirus,
• wherein the cell line does not generate replication competent adenovirus (RCA) and the cell line comprises:
• i) an E1 gene of 2962 bp including both an E1A gene and an E1B gene;
• and
• ii) an E1A gene of 998 bp that is separate from the E1A gene of i) above, an E1B gene of 1808 bp that is separate from the E1B gene of i) above, or both.