AAV pRep/Cap2, GMP-Ready packaging plasmids, providing rep2, cap2 for AAV2 CGMP manufacturing
|In stock||Please Inquire|
|GR-PRC5-KAN||AAV pRep/Cap5, GMP-Ready packaging plasmids, providing rep2, cap5 for AAV5 CGMP manufacturing||In stock||Please Inquire|
|GR-PRC6-KAN||AAV pRep/Cap6, GMP-Ready packaging plasmids, providing rep2, cap6 for AAV6 CGMP manufacturing||In stock||Please Inquire|
* MTA needs to be executed before the plasmids can be propagated
Production Process of AAV pRep/Cap GMP-Ready Plasmids
|Controlled Aspects AAV pRep/Cap GMP-R Plasmid|
|Establishment of E. coli master & working cell banks|
|Full traceability of materials|
|Full room changeover prior to each production|
|Production in segregated and dedicated space|
|Process & change control|
|Aseptic fill/finish and vialing|
|Document support for IND and IMPD filing|
|Master batch records|
|Product release tests|
|AAV pRep/Cap GMP-R Plasmid Release Testing|
|Safety: Endotoxin, sterility|
|Identity: Restriction analysis & DNA sequencing|
|Purity: UV spec, OD260/280|
|DNA homogeneity: Percentage of supercoiled|
|Purity: Residual impurities, host cell DNA, host cell RNA, host cell protein|
|Purity: Cross batch contamination test by Next-Gen sequencing|
AAV pRep/Cap Plasmids Maps
AAV pRep/Cap is Proven For Efficient AAV Packaging
The robust virus packaging productivity of AAV pRep/Cap plasmids has been tested using the triple plasmids transient transfection method. AAV pRep/Cap plasmids are being used in Vigene’s daily AAV production, for both CGMP grade and research grade batches. Please see the productivity data below.
Fig. 1. pRep/Cap plasmids were tested in AAV production in adherent HEK293 cells. Each serotype specific pRC was cotransfected with the AAV transfer vector and pHelper into HEK293T cells for AAV packaging. The productivity data for each AAV serotype is from individual productions with different AAV transfer vectors containing unique genes of interest. AAV viral vectors were purified via IDX gradient ultracentrifugation. The viral genome titers were measured using qPCR with primers targeting the ITRs.
The Diagram of AAV Production Using the Triple Plasmid Transfection Method
Fig. 2. AAV virus production using triple plasmid transient transfection method. AAV viral vectors will be packaged and assembled when the AAV transfer vector, pHelper, and pRep/Cap are cotransfected into HEK293 cells. The AAV transfer vector contains the gene of interest, pHelper plasmid provides Ad helper function genes, and pRep/Cap encodes AAV Rep and Cap.
Need Custom Plasmid Production?
For AAV viral vector packaging, you will also need AAV pHelper plasmid and your pAAV gene-of-interest transfer vector. Vigene has off-the-shelf, premanufactured GMP-Ready pHelper-Kan plasmids, ready for CGMP AAV manufacturing. We can help you with both GMP-Ready and GMP grade plasmid DNA production. Please contact us.