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Gewählte Publikation:

Muralidharan-Chari, V; Wurz, Z; Doyle, F; Henry, M; Diendorfer, A; Tenenbaum, SA; Borth, N; Eveleth, E; Sharfstein, ST.
(2021): PTSelect (TM): A post-transcriptional technology that enables rapid establishment of stable CHO cell lines and surveillance of clonal variation
J BIOTECHNOL. 2021; 325: 360-371. FullText FullText_BOKU

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 (TM) that utilizes an siRNA cloned upstream of the gene of interest (GOI) that is processed to produce functional PTSelect (TM)-siRNAs, which enable cell enrichment. Cells with stably integrated GOI are selected and separated from cells without GOI by transfecting CD4/siRNA mRNA regulated by PTSelect (TM)-siRNAs and exploiting the variable expression of CD4 on the cell surface. This study describes the PTSelect (TM) principle and compares the productivity, doubling time and stability of clones developed by PTSelect (TM) with conventionally developed clones. PTSelect (TM) 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.
Autor*innen der BOKU Wien:
Borth Nicole
Diendorfer Andreas
Henry Miriam
BOKU Gendermonitor:


Find related publications in this database (Keywords)
Cell enrichment
Chinese hamster ovary cells
Clonal variation
Monoclonal antibodies
Recombinant protein production
Silencing RNA


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