Cell Line Generation: Relying on tricks or tools of the trade?

Jeff Jc Hou,Trent P Munro,Kar M Leung,Michael Song,Eric Zhu,Peter P Gray,Martina Jones

doi:10.1186/1753-6561-9-s9-p16

Abstract

Background Cell line generation (CLG) in the scope of bioproduction can be defined as a method to isolate a single cell expressing a recombinant protein of interest. The standard method of CLG often involves the introduction of the transgene into a cell in an attempt to use the cellular machinery for transcription, translation and secretion. The use of random genomic integration via auxotrophic selection markers creates different layers of complexity, which leads to significant variations in growth, productivity and stability among the subsequent population [1]. The use of epigenetic modulators with genetically improved cell lines have improved the “quality” of the resistant pools [2], while high throughput technologies have simplified the clonal isolation process [3]. However this “blackbox” approach still requires the need to screen hundreds or thousands of individual cells to find a line with the right quality attributes for manufacturing [4]. The challenge for CLG is to significantly reduce the timeline of this process while ensuring robustness and quality of the subsequent clones [5].

Highlights

Cell line generation (CLG) in the scope of bioproduction can be defined as a method to isolate a single cell expressing a recombinant protein of interest
By combining the selection with the isolation in a single step, this direct approach allows for a more efficient process in identifying a “serendipity event”, i.e. a single cell that has been transfected with the vector containing the gene of interest, undergone random integration/s at a non-essential locus and has the ability to express the recombinant protein of interest
An initial assessment via the FITC intensity of the in situ fluorescence complex between the Clone Detect (Molecular Devices) and the protein of interest showed a higher signal from the single step method when compared to the standard method

Summary

Introduction

Cell line generation (CLG) in the scope of bioproduction can be defined as a method to isolate a single cell expressing a recombinant protein of interest. The standard method of CLG often involves the introduction of the transgene into a cell in an attempt to use the cellular machinery for transcription, translation and secretion. The use of epigenetic modulators with genetically improved cell lines have improved the “quality” of the resistant pools [2], while high throughput technologies have simplified the clonal isolation process [3]. This “blackbox” approach still requires the need to screen hundreds or thousands of individual cells to find a line with the right quality attributes for manufacturing [4]. The challenge for CLG is to significantly reduce the timeline of this process while ensuring robustness and quality of the subsequent clones [5]

Methods

Results

Conclusion