Isolation of RNA from Field-Grown Jute (<Emphasis Type="Italic">Corchorus capsularis</Emphasis>) Plant in Different Developmental Stages for Effective Downstream Molecular Analysis |
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Authors: | Pradipta Samanta Sanjoy Sadhukhan Subrata Das Alpana Joshi Soumitra K Sen Asitava Basu |
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Institution: | (1) The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD, 4072, Australia;(2) Acyte Biotech Pty Ltd, Brisbane, QLD, 4072, Australia |
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Abstract: | Transient gene expression systems in mammalian cells continue to grow in popularity due to their capacity to produce significant
amounts of recombinant protein in a rapid and scalable manner, without the lengthy time periods and resources required for
stable cell line development. Traditionally, production of recombinant monoclonal antibodies for pre-clinical assessment by
transient expression in CHO cells has been hampered by low titers. In this report, we demonstrate transient monoclonal antibody
titers of 140 mg/l with CHO cells using the episomal-based transient expression system, Epi-CHO. Such titers were achieved by implementing an optimized transfection protocol incorporating mild-hypothermia and through
screening of a variety of chemically defined and serum-free media for their ability to support elevated and prolonged viable
cell densities post-transfection, and in turn, improve recombinant protein yields. Further evidence supporting Epi-CHO’s capacity to enhance transgene expression is provided, where we demonstrate higher transgene mRNA and protein levels
of two monoclonal antibodies and a destabilized enhanced green fluorescent protein with Epi-CHO compared to cell lines deficient in plasmid DNA replication and/or retention post-transfection. The results demonstrate
the Epi-CHO system’s capacity for the rapid production of CHO cell-derived recombinant monoclonal antibodies in serum-free conditions. |
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