High-reactivity cell strain for detecting biologic activity of recombinant human epidermal growth factors
A technology of epidermal growth factor and biological activity, applied in cells modified by the introduction of foreign genetic material, biochemical equipment and methods, biological testing, etc. problem, to achieve the effect of low effective concentration, high accuracy and high application value
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[0059] In the present invention, the Mouse CRISPR Knockout Pooled Library (GeCKO v2) is packaged as a lentiviral library (LentiCRISPR), and the library is used to infect NIH3T3 cells at an MOI value of 0.3, and then the single clones are sorted by flow cytometry and then induced with a low dose of rhEGF. The NIH3T3 cell line with better reactivity to rhEGF was screened out, and the corresponding detection method was established to quantify the activity of rhEGF. The technical process is: screening of NIH3T3 hyperresponsive cell lines (NIH3T3 cells infected with LentiCRISPR and induced with rhEGF) → establishment of rhEGF activity assay method and methodological verification.
[0060] 1. Materials and methods:
[0061] 1.1 Research object: recombinant human epidermal growth factor
[0062] 1.2 Mouse CRISPR Knockout Pooled Library (GeCKO v2) (#1000000052): purchased from Addgene.
[0063] NIH3T3 cells: purchased from ATCC.
[0064] rhEGF standard and samples: kept by the Reco...
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