Evaluation of release specifications for critical raw materials, drug substance and drug product in the production process of recombinant adeno-associated virus vectors
Merikivi-Dahlqvist, Ina Alice (2022)
Merikivi-Dahlqvist, Ina Alice
2022
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi-fe2022050934044
https://urn.fi/URN:NBN:fi-fe2022050934044
Tiivistelmä
In gene therapy, adeno-associated virus (AAV) vectors have shown to be efficient vehicles for delivering the gene of interest to the target site. To date, there are three AAV-based biologicals approved by authorities; Glybera, Luxturna and Zolgensma. The manufacturing process of AAV vectors, a recombinant form of AAV (rAAV), considered in this thesis is based on HEK293 cells. These producer cells are transfected by three plasmids containing rep/cap genes, helper genes and a gene of interest, turning the HEK293 cells into AAV-producing factories. Plasmid delivery into the producer cells is called triple transfection or transient transfection and is achieved using a transfection reagent.
The upstream process that covers the production in producer cells is followed by a downstream process with several purification steps resulting in a purified rAAV drug substance and, after formulation and filling, in an rAAV drug product.
Two critical raw materials in this process are plasmids and transfection reagent. Before these components can be added to the process, they must be qualified to ensure quality, safety, and efficacy. The same considerations apply to the purified drug substance before it can be released for filling and before the drug product can be released for clinical trials or to the market. Medicinal products are supervised and regulated by authorities.
This thesis has collected the regulatory requirements and specifications for plasmids, transfection reagents, and the rAAV vector drug substance and drug product. In addition, associated assays have been defined.
The result of this thesis is a suggestion of tests based on regulatory requirements, the quality by design (QbD) approach, and the reviewed literature, to be included in the specification of plasmids, transfection reagent and the AAV drug substance and drug product.
The upstream process that covers the production in producer cells is followed by a downstream process with several purification steps resulting in a purified rAAV drug substance and, after formulation and filling, in an rAAV drug product.
Two critical raw materials in this process are plasmids and transfection reagent. Before these components can be added to the process, they must be qualified to ensure quality, safety, and efficacy. The same considerations apply to the purified drug substance before it can be released for filling and before the drug product can be released for clinical trials or to the market. Medicinal products are supervised and regulated by authorities.
This thesis has collected the regulatory requirements and specifications for plasmids, transfection reagents, and the rAAV vector drug substance and drug product. In addition, associated assays have been defined.
The result of this thesis is a suggestion of tests based on regulatory requirements, the quality by design (QbD) approach, and the reviewed literature, to be included in the specification of plasmids, transfection reagent and the AAV drug substance and drug product.