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Transient Protein Production in Mammalian Cells
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.
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.