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AssigneeAmsterdam Molecular Therapeutics B.V.
InventorsMasashi Urabe, Keiya Ozawa, Saskia Jacoba Petronella Haast, Wilhelmus Theodorus Johannes Maria Christiaan Hermens
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The present invention relates to the production of adeno-associated viral vectors in insect cells. The insect cells therefore comprise a first nucleotide sequence encoding the adeno-associated virus (AAV) capsid proteins, whereby the initiation codon for translation of the AAV VP1 capsid protein is a non-ATG, suboptimal initiation codon. The insect cell further comprises a second nucleotide sequence comprising at least one AAV inverted terminal repeat (ITR) nucleotide sequence; a third nucleotide sequence comprising a Rep52 or a Rep40 coding sequence operably linked to expression control sequences for expression in an insect cell; and, a fourth nucleotide sequence comprising a Rep78 or a Rep68 coding sequence operably linked to expression control sequences for expression in an insect cell. The invention further relates to adeno-associated viral vectors with an altered ratio of the viral capsid proteins that provides improved infectivity of the viral particles.
A nucleic acid construct comprising a nucleotide sequence encoding adeno-associated virus (AAV) VP1, VP2, and VP3 capsid proteins, wherein the initiation codon for translation of the AAV VP1 capsid protein is CTG, TTG, or GTG and wherein the nucleotide sequence is operably linked to an expression control sequence for expression in an insect cell.
An AAV virion comprising in its genome at least one nucleotide sequence encoding a gene product of interest, which sequence is not a native AAV nucleotide sequence, and wherein VP1 capsid protein is present in the virion in at least 110% of the amount of capsid protein VP2 and at least 11% of the amount of capsid protein VP3, the amount of said capsid proteins being determined using an antibody that recognizes an epitope that is common to each of VP1, VP2 and VP3.
An AAV virion comprising in its genome at least one nucleotide sequence encoding a gene product of interest which sequence is not a native AAV nucleotide sequence, and wherein the virion comprises a VP1 capsid protein comprising leucine or valine at amino acid position 1.
Dependencies are inferred from the claim text.