Stable Cell Lines Development for Research Assay

Biologics International Corp (BIC) provides a stable cell line development service (gene knock-in or knock-out). We can insert any gene of interest into the genome of the cells. Over years of experience in cell culture, we have accumulated comprehensive knowledge of the optimal antibiotic concentrations to use for a variety of stable cells. Stable cell line construction is the best choice for not only the production of recombinant antibodies or recombinant proteins, but also for research in various fields including signal transduction, ion channel research, drug target screening, and immunotherapy drug development. We have successfully cultured dozens of cell lines (CHO, HEK293, HeLa, M14, THP-1, BHK21, HFF-1, HepG2, MCF-7, Vero and more). To obtain a stable single cell clone with target gene inserted, you only need to provide the gene sequence.

Creating stable cell lines is time-consuming and complex. Let us streamline your processes for development of stable cell lines you need for cell-based assays. For further information please do not hesitate to get in touch.

Service Content

Service Name	Deliverables	Timeline	Price
Stable Cell Lines Generation	1 single cell clone (2 tubes/clone) Quality-control report & Experiment process report	~3 months	Inquiry

Key Features

• Rapid delivery: experiments can be performed within 2-3 months and the single cell clones can then be delivered.
• We possess extensive experience in cell culture and are familiar with all kinds of cell culture methods.
• Optimal antibiotic concentrations for common cell lines have already been determined.
• Professional codon optimization technique ensures high gene expression levels.
• Evaluation of transient transfection expression ensures the success of the project.

Service Process for Stable Cell Lines Generation

• Cell preparation: based on the cell type required, we prepare cells 2 weeks in advance to maintain high cell activity.
• Target plasmid construct preparation: based on the specified gene sequence, we perform codon optimization and gene synthesis.
• Preparation:
  • Determine the concentration of antibiotics: screen for cell density and antibiotic concentration. We can determine the minimum concentration needed to achieve 100% cytotoxicity at any time point.
  • Transfection optimization: prepare cells 24 hours before transfection. Perform plasmid extraction and specify transfection conditions.
• Positive clone screening:
• Transfer the transfected cells to 96-well plates and screen the cells using selective medium based on limiting dilution analysis.
• Plating density determination: 100-1000 cells/wel.l
• Confirmation and amplification of positive clones:
• After 3-4 weeks, mark up the single clones and check the titers from the supernatants using ELISA.
• Screen and scale up the positive clones.
• Stability confirmation:
• Cells with the same concentration are inoculated into a 6-well plate. We count the cell numbers every 48 hours. Finally, we confirm the expression stability of the cells using ELISA/WB assay.
• Stability test for 10 passages.
• Mycoplasma detection:
• Q-PCR and Western Blot.
• Frozen and delivery of the stable cell lines.

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