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PTSelect™: A post-transcriptional technology that enables rapid establishment of stable CHO cell lines and surveillance of clonal variation
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  • Vandhana Muralidharan-Chari,
  • Zachary Wurz,
  • Francis Doyle,
  • Matthew Henry,
  • Andreas Diendorfer,
  • Scott Tenenbaum,
  • Nicole Borth,
  • Edward Eveleth,
  • Susan T. Sharfstein
Vandhana Muralidharan-Chari
SUNY Polytechnic Institute
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Zachary Wurz
Hocus Locus
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Francis Doyle
SUNY Polytechnic Institute
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Matthew Henry
The University of Queensland
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Andreas Diendorfer
University of Natural Resources and Life Sciences
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Scott Tenenbaum
SUNY Polytechnic Institute
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Nicole Borth
University for Applied Live Sciences
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Edward Eveleth
Hocus Locus
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Susan T. Sharfstein
SUNY Polytechnic Institute, SUNY Polytechnic Institute
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Abstract

Currently, stable Chinese hamster ovary cell lines producing therapeutic, recombinant proteins are established either by antibiotic and/or metabolic selection. Here we report a novel technology, PTSelect™ that utilizes an siRNA cloned upstream of the gene of interest (GOI) that is processed to produce functional PTSelect™-siRNAs, which enable cell selection. Cells with stably integrated GOI are selected and separated from cells without GOI by transfecting CD4/siRNA mRNA regulated by PTSelect™-siRNAs and exploiting the variable expression of CD4 on the cell surface. This study describes the PTSelect™ principle and compares the productivity, doubling time and stability of clones developed by PTSelect™ with conventionally developed clones. PTSelect™ rapidly established a pool population with comparable stability and productivity to pools generated by traditional methods and can further be used to easily monitor productivity changes due to clonal drift, identifying individual cells with reduced productivity.

Peer review status:UNDER REVIEW

20 Apr 2020Submitted to Biotechnology and Bioengineering
21 Apr 2020Assigned to Editor
21 Apr 2020Submission Checks Completed
26 Apr 2020Reviewer(s) Assigned
12 May 2020Review(s) Completed, Editorial Evaluation Pending