	US 12,448,628 B2
	Further improved AAV vectors produced in insect cells
Jacek Lubelski, Amsterdam (NL); Sebastiaan Bosma, Amsterdam (NL); Harald Peter Albert Petry, Amsterdam (NL); and Wilhelmus Theodorus Johannes Maria Christ Hermens, Amsterdam (NL)
Assigned to Uniqure IP B.V., Amsterdam (NL)
Filed by Uniqure IP B.V., Amsterdam (NL)
Filed on Nov. 5, 2020, as Appl. No. 17/090,807.
Application 17/090,807 is a division of application No. 15/124,139, granted, now 10,837,027, previously published as PCT/NL2015/050149, filed on Mar. 10, 2015.
Claims priority of application No. 14158610 (EP), filed on Mar. 10, 2014.
Prior Publication US 2021/0222198 A1, Jul. 22, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/86 (2006.01); C07K 14/005 (2006.01); C12N 7/00 (2006.01)

CPC C12N 15/86 (2013.01) [C07K 14/005 (2013.01); C12N 7/00 (2013.01); C12N 2710/14043 (2013.01); C12N 2750/14121 (2013.01); C12N 2750/14122 (2013.01); C12N 2750/14143 (2013.01); C12N 2750/14152 (2013.01); C12N 2799/026 (2013.01); C12N 2830/008 (2013.01)]

20 Claims

1. A method for producing an AAV in an insect cell, comprising:

(a) culturing an insect cell comprising a nucleic acid construct comprising a nucleic acid molecule having a nucleotide sequence comprising an open reading frame, wherein the reading frame in 5′ to 3′ order consists of:

(i) a first codon, which is a suboptimal translation initiation codon selected from the group consisting of CTG and ACG;

(ii) a second codon encoding an amino acid residue selected from the group consisting of alanine, glycine, valine, aspartic acid and glutamic acid; and

(iii) a sequence encoding wild-type adeno-associated virus (AAV) serotype 5 capsid proteins, wherein the sequence lacks only the wild-type VP1 translation initiation codon,

(b) recovering of the AAV.