High throughput screening identifies novel, cell cycle-arresting small molecule enhancers of transient protein expression
| Autor: | Shirley Ng, Dorothea Reilly, Pamela Chan, Mark Zak, Athena W. Wong, Rebecca Turincio, Juan Li, Hermann-Josef Meyer, Blair Wilson, Maureen Beresini |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine High-throughput screening CHO Cells Biology Transfection 01 natural sciences Antibodies law.invention Small Molecule Libraries 03 medical and health sciences Cricetulus law Cricetinae 010608 biotechnology Gene expression Animals Humans Polyethyleneimine HEK 293 cells Antibody titer Cell Cycle Checkpoints Cell cycle Molecular biology Small molecule Recombinant Proteins High-Throughput Screening Assays Titer HEK293 Cells 030104 developmental biology Immunoglobulin G Antibody Formation Recombinant DNA Biotechnology |
| Zdroj: | Biotechnology Progress. 33:1579-1588 |
| ISSN: | 8756-7938 |
| Popis: | Transient gene expression in mammalian cells is an efficient process for producing recombinant proteins for various research applications to support large molecule therapeutics development. For the first time, we report a high throughput small molecule (SM) screen to identify novel compounds that increase antibody titers after polyethylenimine (PEI) transient transfection of a HEK293 cell line. After screening 31,413 SMs in a 50 μL scaled-down process, we validated 164 SMs to improve yields by up to twofold. The titer increase mediated by the SMs varied for different antibodies. SM dose optimizations resulted in almost threefold higher titers. The top 2, structurally distinct SM hits, increased antibody titers more than twofold in a 1 mL production process. Averaged across three antibodies of different expression levels, the compounds enhanced transient productivity by ∼80%. Intriguingly, both compounds arrested cells in the G2/M cell cycle phase leading to a decrease in growth and nutrient consumption, while elevating titer, nuclear plasmid DNA (pDNA) copy numbers, and mRNA levels. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 3:1579-1588, 2017. |
| Databáze: | OpenAIRE |
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