Hi All,
Does anyone have a good protocol for transient transfection of mammalian cells using separate plasmids for heavy and light chain of a recombinant antibody? What range of ratios should I try for LC:HC?
Hi Alden,
Unfortunately, I cannot help you with transiently transfecting plasmids (because normally the good thing with plasmids is, that they are transfected for durable overexpression). If you want to transiently express your heavy and light chains I can recommend transcribing your plasmids into mRNA and then transfecting the mRNA in your cells. We developed a protocol for that (transfection works for peritoneal and bone marrow macrophages and for MEFs). If you want, you can have a look at :
Herb, M., Farid, A., Gluschko, A., Krönke, M., Schramm, M. Highly Efficient Transfection of Primary Macrophages with In Vitro Transcribed mRNA. J. Vis. Exp.
I hope that helps you.
All the best,
Marc
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