Applications
Products
Service
Company
Seattle Genova is here and ready to assist you. The Stable Cell Line Services offered by Seattle Genova are prepared to meet all of your custom cell line requirements, including those for reporter cells that express GFP, luciferase, or other enzymes for high-throughput assays, overexpression cell line construction, or uniquely engineered cells for any research purpose. We have access
to a wide array of technologies and a skilled team with experience with a range of cell line technologies. You will receive our top-notch customer service, which includes regular communication, quick turnaround times, and high success rates.
Our stable cell line generation service area
1. Overexpressing cell line Construction
-Overexpressing your gene of interest (GOI) generation by lentiviral transduction then validated by RT-qPCR.
-Also can be validated using Western blot and immunohistochemistry upon request.
2. Gene knock-out cell line Construction (CRISPR Technology)
- A pair of gRNAs targeting the GOI is introduced into the target cells along with Cas9.
- Sanger sequencing and the final product will undergo sterility test and mycoplasma detection as a standard QC procedure.
3. Luciferase tagging cell line Construction
- Firefly luciferase substrate which produces strong and stable light signal.
- Routine mycoplasma testing.
4. GFP tagging cell line Construction
- The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9kDa) that exhibits bright green fluorescence when exposed to
light in the blue to ultraviolet range.
- Fluorescent tracing can be done on HEK293 and MCF-7 cells or other cell lines upon request.
5. CHO stable cell line for protein production
- Using our self-developed CHO cells and adopt GS/DHFR expression system
- Single clone screening and Mycoplasma QC testing.
6. shRNA knockdown stable cell lines
- Three lentiviruses will be transduced carrying the top 3 shRNAs, which the best knockdown efficiency validated by RT-qPCR will be delivered
- Further QC testing of mycoplasma.
7. Tet inducible gene expression stable cell lines
-Generate Tet regulatory protein expression stable cell lines (e.g. rtTA, tTS/rtTA, etc.)
-Validated by RT-qPCR and further QC testing of Mycoplasma
8. Customized stable cell line generation
-Point mutation, gene knock-in, etc. (upon request)
Our stable cell line workflow
●Cell preparation
We have successfully created a variety of cell lines, such as CHO, and cells will be prepared two weeks in advance to ensure a highly active state during transfection. At the same time, we also accept cells given by our customers.
●Determination of antibiotic screening concentration
We have decades of experience and are aware of the best screening concentration of various types of antibiotics.
●Plasmid construction and cell transfection
●Screening, detection and amplification of positive clones
The transfected cells are plated onto a 96-well plate utilizing a limited dilution method and the clones are screened using a specific screening medium. Mark up the single clones/cell pools and examine the ELISA titers from the supernatants. After that, Screen and scale up the positive clones.
●Pressure selection
To expand the protein expression level of cell lines, a pressure selection is needed to carry out the construction of cell lines for protein production.
●Subcloning and single clone screening
Select the clone/pool with the biggest expression and perform monoclonal screening by limited dilution. After 3-4 weeks, we will mark up the single clones and examine the ELISA titers from the supernatants and scale up the positive single cell clones.
●Quality control of cell lines
Stability test: Cells of the same density were inoculated into flasks and tested every 48 hours for counting. Then cells were continuously inoculated with a similar initial density and the expression stability is determined by ELISA.
Identify and freeze stable producers: Based on the results of growth and production profile, we will select the top 5-10 clones to be frozen.
The highlights if our service
Our cell line construction services are aligned with your priorities in mind for any molecular format. The processes used to generate your cell line are designed to provide high product titers and the necessary product qualities. To develop master and working cell banks for further manufacturing application, our licenced manufacturing facility uses a specialised cGMP unit. The cell banks are characterized in accordance with FDA and European regulatory agency requirements.
Deliverable
Two vials (>1 x 106 cells/vial) of Polyclonal or monoclonal cell populations
QC data that includes Western Blot and/or qPCR data (sequencing and flow cytometry also available)
Glycerol stock, plasmid DNA, and/or lentiviral-vector clone
Stability testing is available
Milestone reports and detailed final report
Contact Us Now!
Please contact us with your project demands if you don't find the cell line service you're looking for listed here. Our scientific team can provide tailored cell lines for any purpose. Additionally, you may email us at info@seattle-genova.com.
Thank you for your interest in our Stable Cell Line Construction Services services. Please complete the form below and we will contact you shortly.
Fields indicated by an * are required.
