Surface display vectors for selective detection and isolation of high level antibody producing cells.

Cell line generation for production of biopharmaceuticals in mammalian cells usually involves intensive screening of clones to identify the rare high producers. In order to facilitate efficient and selective fluorescence activated cell sorting (FACS) based enrichment and cloning of antibody producing CHO cells, we developed a special vector setup by inserting a leaky translation termination signal between the heavy chain of an IgG antibody and an IgG transmembrane domain. Partial read-through during translation of the antibody heavy chain leads to display of a subset of the produced antibody on the surface of the expressing cell.

Mammalian stable expression of biotherapeutics.

Many therapeutically relevant proteins, like IgG antibodies, are highly complex, multimeric glycoproteins that are difficult to express in microbial systems and thus usually produced in mammalian host cells. During the past two decades, stable mammalian expression technologies have made huge progress resulting in highly increased speed of cell line development and yield of manufacturing processes. Here, we give an overview of technologies that are applied at different stages of state-of-the-art cell line development processes for biomanufacturing.

Combination of the 2A/furin technology with an animal component free cell line development platform process.

The recently described 2A/furin technology combines both chains of the antibody in a single open reading frame. Upon translation and secretion, the peptide is processed by the cell to generate native fully functional IgG antibodies. Here, we describe the results of an evaluation study of this technology for an industrial CHO cell line development process.