Production of Lentiviral Vectors
A high-volume gene therapy vector manufacturing process which produces a recombinant gene therapy vector which is able to transform host cells even when they are not dividing.
1 . A method comprising:
a. Transducing a mammalian cell with a baculovirus to make a transduced mammalian producer cell; and then
b. Culturing said transduced mammalian producer cell in culture media, and harvesting from said transduced mammalian producer cell and/or culture media a second virus having a therapeutic transgene.
2 . The method of claim 1 , wherein said harvesting comprises harvesting at about 144 hours after said transduction.
3 . The method of claim 1 , wherein said second virus is derived from a virus which in its wild state has a single-stranded genome.
4 . The method of claim 1 , wherein said second virus is able to transduce a human cell which is not actively dividing.
5 . The method of claim 1 , wherein said second virus is replication deficient.
6 . The method of claim 1 , wherein said second virus can integrate in host genome at specific site and can produce stable long-term expression.
7 . The method of claim 1 , wherein said second virus is non-immunogenic.
8 . A recombinant baculovirus having at least one nucleic acid sequence coding for a second virus derived from a virus which in its wild state has a single stranded genome, said second virus being replication-deficient and having a therapeutic transgene.
9 . The recombinant baculovirus of claim 8 , wherein said second virus is able to transduce a human cell which is not actively dividing.
10 . The recombinant baculovirus of claim 8 , wherein said second virus can integrate in the human genome and can produce stable long-term expression in a transduced human cell.
11 . A method comprising:
a. Obtaining the recombinant baculovirus of claim 8 ; and then
b. Transducing a mammalian producer cell with said recombinant baculovirus to make a transduced mammalian producer cell; and then
c. Culturing said transduced mammalian producer cell in culture media, and harvesting said second virus from said transduced mammalian producer cell and/or culture media.
12 . A viral vector able to transfect a human cell which is not actively dividing, said viral vector produced by a mammalian producer cell transduced with a recombinant baculovirus.
13 . The viral vector of claim 12 , wherein said viral vector has a therapeutic transgene.
14 . The viral vector of claim 12 , said viral vector comprises virus derived from a virus which in its wild state has a single stranded genome.
15 . A method comprising:
a. Obtaining the viral vector of claim 12 ; and
b. Transducing a human patient's cells with said viral vector.
16 . A method comprising:
a. Obtaining the viral vector of claim 13 ; and
b. Transducing a human patient's cells with said viral vector.
17 . A method comprising:
a. Obtaining the viral vector of claim 14 ; and
b. Transducing a human patient's cells with said viral vector.