A screening method for gs expression system cell line
An expression system and screening method technology, applied in the field of high-throughput screening of GS expression system cell lines, can solve the problems of long experimental period, heavy workload, cumbersome operation, etc., and achieve shortened screening period, high clone formation rate and stable sex-enhancing effect
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Embodiment 1
[0031] Example 1 Preparation of cell pool A and monoclonal screening of cell pool A for stable cell lines with high expression
[0032] 1. Construction of expression constructs
[0033] Referring to WO 2007 / 014162 A2 patent and GS Xceed TM The Gene Expression System (purchased from Lonza Company) was used to independently prepare the Anti-VEGF humanized antibody GS double gene expression vector.
[0034] 2. Transfection of Mammalian Cells
[0035] use HiPure Plasmid DNA Purification Kit (Invitrogen, K2100-04) was used to extract the double-gene expression vector of Anti-VEGF humanized antibody, linearized and purified by PvuI (TaKaRa, 1242A), and the total amount of 40 μg DNA / electroporation system was used CHOK1SV GS-KO cells (Lonza, GS Xceed TM GeneExpression System), resuspend the cells after three electric shocks in a 125mL conical culture flask (corning, 431143-50EA) containing 30mL CD CHO Medium (Invitrogen, 12490-25), 140rmp, 37°C, 8.0% CO 2 Shaking culture under c...
Embodiment 2
[0073] Example 2 Preparation of Cell Pool B and Cell Pool B Monoclonal Screening for High Expression Stable Cell Lines
[0074] 1. Construction of expression constructs
[0075] Referring to WO 2007 / 014162A2 patent and GS Xceed TM The Gene Expression System (purchased from Lonza Company) was used to independently prepare the Anti-VEGF humanized antibody GS double gene expression vector.
[0076] 2. Transfection of Mammalian Cells
[0077] use HiPure Plasmid DNA Purification Kit (Invitrogen, K2100-04) was used to extract the double-gene expression vector of Anti-VEGF humanized antibody, linearized and purified by PvuI (TaKaRa, 1242A), and the total amount of 40 μg DNA / electroporation system was used CHOK1SV GS-KO cells (Lonza, GS Xceed TM GeneExpression System), resuspend the cells after three electric shocks in a 125mL conical culture flask (corning, 431143-50EA) containing 30mL CD CHO Medium (Invitrogen, 12490-25), 140rmp, 37°C, 8.0% CO 2 Shaking culture under condition...
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