Work package 3
Optimization, Validation and Definition of Release Criteria of the Gene Therapy Medicinal Product
Work package 3 deals with the assessment, the optimisation and the validation of the quality and safety profile for each component, implemented in the manufacturing process of the Gene Therapy Medicinal Product (GTMP) as well as the final patients’ surgery procedure.
These components are realised by partners (Fig. 1) as following:
|Iris (IPE) and retinal pigment epithelial (RPE) cells extracted in analogy to patient`s autologous pigment epithelial cells from
|The plasmid components applied via electroporation transfection technology
|The transfection equipment
Prior to the animal testing it was demonstrated by in vitro experiments that freshly isolated IPE as well as RPE cells can be transfected with the Sleeping Beauty transposon system with high efficiency and cell viability. Transfection efficiency and long-term protein expression (proofed via Western Blot analysis, Fig. 2) as well as immunohistochemistry and gene expression analysis were used to analyse morphological and genomic stability. Additionally, quantity and quality of the isolated cells were evaluated by microscopy, standardised cell viability assays and cell culture analysis. The consistency of the final product in adequate low cell amounts to the prospective patients’ biopsy is a key element of this WP.
The electroporation technique, a standard technique in gene delivery, is evaluated in relation to transfection efficiency and cell compatibility. An electroporation device with cell holding cuvettes, electrodes and buffers will be adapted to the electroporation procedure of the required low cell amount (5’000 to 10’000 cells) yielding by the patient`s biopsies. The optimal transfection conditions are used for the transfection of the primary human IPE cells.
The validation of defining release criteria and in-process testing will be an important guideline for the training of the personnel responsible for GTMP (Gene Therapeutic Medicinal Product) production during the clinical trial in Switzerland.