Characterization of the Novel Human AGE1hn Cell Line for Production of Recombinant Proteins

In this study the process suitability of AGE1hn, a new human production cell line for biologicals developed at ProBioGen was assessed. This permanent cell line was generated by transfection of cells originating from the periventricular zone of foetal human brain with Adenovirus 5 E1A and B genes, a combination of genes with a high safety profile. The E1 genes are responsible for immortalisation and are stably maintained in the absence of selection pressure. Following adaptation to growth in suspension in serum-free media the cell line was further modified to express therapeutic glycoproteins or modulator proteins enhancing the yield of attenuated viruses. To characterize the cell line metabolically and to determine its synthetic capacity alpha-1-antitrypsin was chosen as model protein. This 52 kDa protein has three N-linked glycosylation sites and belongs to the serpine family. Cultivation in several different systems (e.g. T-Flasks, shaker flasks, paddle agitated vessels and stirred tank reactors), with varying process strategies and cultivation parameters was carried out. Because of growth inhibition due to cell-cell contact observed in the AGE1hn cell line, a substantial agitation was shown to be necessary in all cultivation systems. Moreover, the metabolic profile of the cells was determined, revealing differences to other commonly used cell lines (e.g. CHO cells).