Transient Protein Production in Mammalian Cells

By on April 13, 2015
Ask the Expert

This Ask the Expert will address questions in regards to transient protein production in mammalian cells.  Mammalian transient expression allows for the rapid generation, purification, and characterization of milligram to gram quantities of secreted or intracellular recombinant proteins for therapeutic, functional, and structural studies.

Questions may include:

  • Getting started with Mammalian Transient Expression Systems
  • Key Elements Necessary for the Establishment of a Mammalian Transient Production System
  • Scaling Transient Protein Production to Accommodate a Wide Variety of Early Discovery Studies and Applications
  • Optimizing the Transient Protein Production Process
  • Tools and Strategies for Purification and Evaluation of Transiently Expressed Proteins

Who Should Attend?

Scientists interested in setting up a rapid and robust mammalian expression process to produce milligram to gram quantities of recombinant protein.

This Ask the Expert session is sponsored by Thermo Fisher Scientific and hosted by Jonathan Zmuda, Ph.D. Dr. Zmuda is the Associate Director of Cell Biology in the Life Sciences Solutions Group at Thermo Fisher Scientific, Inc.  Prominent among his roles, Dr. Zmuda leads a team dedicated to discovering and developing new technologies and products useful for cell biology applications including protein expression, cell culture, rare cell analysis and instrumentation while also focusing on the development, qualification and validation of cell-based assays used for QC testing.  Dr. Zmuda received his Ph.D. in Cell Biology from the University of Maryland, College Park and his undergraduate degree from Dickinson College in Carlisle, PA.

Tips:

Best practices for maintaining your cells before, during and after transfection:

Cells used for transient protein expression should be well characterized in terms of their growth rates/doubling times, growth profiles to maximum viable cell density and passage stability over time.  From the time of thaw, care should be taken to ensure that cells are routinely sub-cultured when cells reach approximately 1/3 to 1Ž2 of their maximal viable cell density; cells should not be over-split before they reach log phase growth, neither should they be allowed to grow to exceedingly high densities before sub-culturing.  Viability at the time of transfection should be high (~95% or greater) and viability and viable cell density should be monitored post-transfection to understand these parameters in your system.

Ensuring optimal complexation of your DNA and transfection reagent:

All transfection reagents are different from one another, thus there is no one best method for optimal transfection complexation.   During optimization, volumes of DNA and transfection reagents should be determined empirically, as well as your choice of complexation medium, complexation temperatures and times. 

Please take advantage of the opportunity to ask our expert a question and participate in a lively discussion on transient protein production in mammalian cells!  We will also be posting tips, so check in regularly.

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