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•Laser Enabled Analysis And Processing Data Supports Superior Performance Of Animal Component Free Electroporation And Recovery Formulation

Electroporation is widely used for gene delivery in cell line generation, and is the method of choice for serum-free suspension culture transfection. Electroporation protocols and media can have a major impact on post-electroporation cell viability, recovery cell density, and transfection efficiency.

SAFC Bioscience's animal component free (ACF) electroporation and recovery formulation was previously shown to achieve equivalent or higher post-electroporation viabilities as well as equivalent or higher viable cell densities after 24 hours recovery compared to other media tested. These results were confirmed. Using a fluorescence platereader to measure transfection level, this formulation was also shown to facilitate higher transient protein expression levels than other media. Recently, we were able to more accurately measure transfection levels in different media conditions using Laser Enabled Analysis and Processing (LEAP).

LEAP is a technology that can be utilized for a number of purposes including transfection of macromolecules, assessment of cell viability and growth, assessment of expression of secreted and non-secreted proteins by individual cells, and targeting and destruction of selected cells in a population. For our purposes, the LEAP instrument was used in two ways. First, we used it to calculate the number of cells expressing transfected plasmid DNA compared to the total number of cells present for each test condition. This value was labeled transfection efficiency. Second, we used it to quantify the level of Green Fluorescent Protein (GFP) expression per transfected cell in the samples analyzed. Our results indicated that cells electroporated and recovered in our optimized formulation have an equivalent transfection efficiency compared to competitor media tested. However, cells transfected and recovered in our formulation exhibit a population shift that reflects higher levels of recombinant protein expression per transfected cell.

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•Laser Enabled Analysis And Processing Data Supports Superior Performance Of Animal Component Free Electroporation And Recovery Formulation

SOURCE: SAFC