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For Assay Development Stable Cell Lines Service, please click here.

GenScript Production Stable Cell Line Service Options

• Host cells: CHO, CHO-K1, Glutamine Synthetase-knockout CHO-K1, HEK293, 293T, 293EC18, 293EC18/CRE-Luc, 1321N1, RH7777, U-2OS, or customer-specified cell line.
• Genes: Antibodies, GPCRs, ion channels, proteases, kinases, transmembrane proteins and more.
• Customized Service: Choose part or all of the following steps based on your specific needs.

Stable Cell Line Service Details

• Service Description
• Service Specifications

Steps	Gene synthesis & subcloning (optional)	Transient expression evaluation	Stable cell line development	Process development (included with SC1557)
Deliverables	Target gene(s) in pUC57 & vector map	Expression report	Top 3 stable clones (5 clones in case of SC1557)	Comprehensive report
Description	GenScript synthesizes codon-optimized gene Subcloning into mammalian cell expression vector	Transfect cells with expression constructs using small scale expression system Detect expression levels by ELISA and/or WB	Stable transfection and screening MTX amplification and screening (SC1503 & SC1557) Subcloning of top 3 clones Screening subclones with highest productivity Stability test for 10 passages Batch culture to evaluate final expression Research cell bank preparation	Medium selection Fed-batch process development –Physical compatibility –Feed supplementation test

		Guaranteed Gram Level (SC1709)	Stable Bioproduction (SC1503)	Advanced Bioproduction (SC1557)
Features	Selection system	Glutamine Synthetase or DHFR
	Guaranteed yield	V	-	V
Steps	Gene synthesis & subcloning (optional)	V	V	V
	Transient expression evaluation	V	V	V
	Stable cell line development	V	V	V
	Process development	optional	optional	V
	Production service	optional	optional	optional
Deliverables	Target gene(s) in pUC57 & vector map	V	V	V
	Expression Report	V	V	V
	Top 3 stable clones (5 clones for SC1557)	V	V	V
	Comprehensive report	-	-	V
Turnaround time	weeks	14-17	26-29	36-39

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We offer GLP compliant services with all 3 packages.

Flow Chart Of GenScript Stable Transfection & Cell Line Development Service

High quality cell lines start with high quality DNA

Our customers have the option of providing their own DNA for stable cell line generation purposes, however, GenScript recommends gene synthesis to ensure that we use high quality, codon optimized genes for the best expression levels.

Stable transfection

GenScript allows you to choose from either Glutamine Synthetase or DHFR gene expression systems to construct your stable cell line. To start your project, DNA carrying the gene of interest must be introduced into the cell with a stable transfection protocol. Depending on your preference, we will use either plasmid-based stable transfection or lentivirus-based infection as your preferred stable transfection method.

Transient expression evaluation

To ensure that your protein of interest is expressed in the cells, we first characterize your protein expression by ELISA or Western Blot 48 hours after the transfection.

Stable cell line development

Transfected cells are grown in the appropriate selective media to generate stable pools. For packages using a DHFR expression system, we can also subject the stable pools to additional Methotrexate (MTX) amplification and screening, in order to increase expression levels (packages using a Glutamine Synthetase expression system do not require gene amplification). The productivity and quality of the pools is evaluated up to 10 passages for the top 10 clones.

Clone picking

Top 3 clones from each pool are picked and evaluated further, based on their growth rates.

Cell line stability screening

Stability testing is a critical component of characterizing bioproduction cell lines. Stable cell lines in continuous culture suffer from genetic instability as the number of passages increases. Stability test will help you ensure the quality of your production cells over the intended passages.

Final deliverables, QC data and full report

We provide the sequence and construct report (in case of synthesized gene), top 3 stable clones and a comprehensive stable cell line generation report