The overall goal of the project is to demonstrate the technical viability of a process for the production of a heterodimeric fusion protein at pilot scale. The expression system, as well as the fusion protein produced, at laboratory scale fulfills the specifications needed. Past experience has shown that it is not possible to predict whether this will be the case after upscaling. It will be necessary to address this in order to fulfill the material demand for preclinical and clinical evaluation. The objectives of the project are: - process definition of an established laboratory scale technology using a new expression system for a new product at pilot scale
- provision of material for the preclinical evaluation of the therapeutic concept
- consideration of regulatory requirements necessary for clinical application of the product.
- presentation of the results to an extended audience such as the Animal Cell Technology Industrial Platform (ACTIP), regulatory experts and clinicians. - exchange of technology between partners to accelerate product development. This proposal combines the experience of some very skilled European groups in molecular biology (Dr. Hauser, GBF), cell culture engineering (Prof. Carrondo, IBET; Prof. Griffiths, CAMR; Dr. van Weperen, Bio-Intermediair), down-stream processing (MERCK), glycoprotein chemistry (Dr. Conradt, GBF) and pharmaceutical development (MERCK) to realise a unique demonstration project: In an ongoing project the partners have established cell lines, analytical procedures and fermentation systems for protein production at laboratory scale. In the proposed project the partners will compare different cell culture techniques at pilot scale using the minimal protein medium formulation, as used at laboratory scale, to identify the most reproducible and economic production process for industrial application. Product consistency will be closely monitored by state of the art analytical methods.
The product is a heterodimeric fusion protein consisting of a monoclonal antibody and the cytokine TNFoc which is developed at MERCK for the novel concept of "targeted tumor therapy". The partners have demonstrated, for the first time, production of the fusion protein in an expression system which advances from current state of the art expression systems by: - an unusual expression unit driven by a single strong promotor hybrid which contains three protein chain-encoding genes which are cotranslated from one tricistronic mRNA molecule. This guarantees strict coupling of expression and allows stoichiometric production of IgG light and heavy-cytokine chains. - genetic constructs established in a preselected BHK-21 sub-cloned which show hi levels of expression of foreign genes and a defined protein glycosylation pattern.
MERCK as an European company is very active in the development of biologics for tumor therapy and this project will accelerate the establishment of biotechnology in the company and facilitate the realization of future biotechnology projects.
Through the project all the partners will participate in the knowledge of the entire group. This experience will facilitate the realization of future projects in this field.
Funding SchemeCSC - Cost-sharing contracts
9700 AL Groningen
SP4 0JG Salisbury