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Standard Services of Stable Cell Line Development
Note: Price/availability/specifications subject to change without notice. If not specified, all products and services are for in vitro research use only. Not for use in or on humans or animals.
Order offline: Phone: 1-617-401-8149, Fax: 1-617-606-5019, Email: message@sydlabs.com, or leave a message with a formal purchase order (PO) or credit card.
Features:
One-stop gene synthesis and DNA subcloning.
Various mammalian expression vectors developed by ourselves or our partners.
Free sequence optimization to meet your goals such as protein expression at high and reliable levels. DNA constructs of any sequence up to 12 kb in length.
We may synthesize codon-optimized cDNA, gene variants, artificially designed DNA, or any other sequence for your research.
Rapid turnaround of stable cell line development: 6-7 weeks.
We have a lot of experience in dealing with difficult-to-express proteins.
While the success rate of stable cell line generation is highly dependent on the target genes and Syd Labs can not guarantee 100% success rate, our in-house data show a success rate greater than 90%.
Syd Labs offers custom services for stable cell line generation for single gene of interest under the constitutively active EF1C promoter. This service includes the subcloning of clients' genes-of-interest into our proprietary mammalian expression vector pLEV343 with zeocin selectable marker.
Stable cell lines are engineered using Zeocin Enhanced technology, with our proprietary multi-cistronic mammalian expression vector pLEV440, to simultaneously express two individual genes.
High expressor stable cell line are developed with patented RUM (Rapid Universal Selectable Marker) cell line creation technology.
Description:
We provide the services of developing stable cell lines for: high production of biomolecules such as monoclonal antibodies and recombinant proteins; G-Protein Coupled Receptors (GPCR) of high pharmacological relevance; other cell-based assays such as ion channel and kinases (RTKs). Our high-throughput cell culture capacity and expertise in medium design and cell adaptation to serum-free suspension growth enable us to offer high quality services at significant cost saving to our clients.
1. Gene synthesis and subcloning:
Subclone the synthesized DNA fragment into the mammalian expression vector commercially available or developed by customers, ourselves or our partners. Cell line stability and clone expression may be improved using our expression vectors. We normally use expression vectors with the zeocin selectable marker. Any DNA sequence of your interest will be synthesized with sequence optimization to meet your goals such as protein expression at high and reliable levels. With an integrated system combining gene synthesis and PCR cloning, we can produce DNA constructs of any sequence up to 12 kb in length. We may synthesize codon-optimized cDNA, gene variants, artificially designed DNA, or any other sequence for your research.
Deliverables:
a. 4-10 µg lyophilized DNA cloned in a plasmid with sequence optimization, synthesis, cloning, verification by sequencing and restriction digestion;
b. E. coli strains containing the plasmids with the desired sequence;
c. Plasmid map and DNA sequence.
2. Transient expression evaluation:
Transfection into mammalian cells and analysis of the short term impact of altered protein expression by ELISA or Western Blot. We normally use monolayer CHO cells and expression vector with the zeocin selectable marker.
Deliverables:
Transient expression evaluation report.
3. Stable cell line development using mammalian cells:
Transfection into mammalian cells and screening for cell lines permanently expressing specific genes through their incorporation into the cellular genome. The standard stable cell line generation service by default uses monolayer CHO cells and expression vector with the zeocin selectable marker. Clones will be screened using ELISA, Western, or cell-based assay of your interest. Our faster, more predictive screening techniques may significantly increase speed of stable cell line development while improving quality and uniformity.
Deliverables:
a. 10 individual clones, 2-5 million cells per vial per clone;
b. Stable cell line development report.
4. High expressor stable cell line development:
Compared to cell lines harboring only one copy of target gene, the cell lines harboring multiple copies of the same target genes are likely to express the target gene at higher levels, which is desirable in many downstream applications. We have made several innovative advancements in its vector technology and stable cell line selection systems that enable not only rapid stable cell line generation but also selection of stable cell lines that contain multiple copies of the target gene cassette. We use a variety of gene amplification-compatible selectable markers, including a new proprietary selectable marker we developed. This new selectable marker has radically improved stable cell line generation process, and has achieved significantly increased the success rate of stable cell line development for difficult targets. The expression levels and stability of target genes can now be tightly controlled and monitored.
Deliverables:
a. 10 individual clones, 2-5 million cells per vial per clone;
b. Stable cell line development report.
Related Links:
- DNA Subcloning Service
- Custom Recombinant Antibody Production Services
- Protein Expression in Mammalian Cells
- Cloning Heavy- and Light-chain Variable Regions of Hybridoma Cell Lines
- Antibody cloning and sequencing service for $1500 only
