Home » Therapeutic Antibody » Antibody Engineering » In vitro Pharmacology » Overexpression Cell Line Construction

Conventional Cell Line Engineering Service

Stable cell lines are used in a variety of applications, including gene functional studies, binding/blocking screening as well as cross-species binding screening during antibody drug development. Generation of a stable cell line refers to the process of the integration of a foreign gene into the host cell chromosome, thus demonstrating expression of a transfected gene insert.

GenScript ProBio possesses a series of stable cell line construction technologies and is experienced in mammalian cell culture and gene-editing experiences. We are capable of constructing a variety of stable cell lines and functional reporter assay cell line through CRISPR, Tet-On technology, lentivirus infection, electroporation, and lipo-transfection, thus shortening your project turnaround time.

Please download the project evaluation form, fill it and send it to us for an evaluation

Major Challenges for Stable Cell Line Generation

Major Challenges Resolution
Lengthy development hindering the downstream applications One-stop service: gene synthesis, plasmid construction and preparation, lentivirus packaging in one service package.
Low efficiency in gene delivery to cells such as suspension cells and primary cells using conventional transfection methods

Lentivirus infection: Lentivirus can infect suspension cells and primary cells.

Non-lentivirus infection: Electro-transfection and nuclear-transfection are utilized for lentivirus-sensitive cell lines

Difficulty in delivery of genes longer than 3 kb into the target cells Lentivirus, electro-transfection and nuclear-transfection are great methods for improved transfection efficiency

Stable Cell Lines at GenScript ProBio

GenScript ProBio has successfully constructed more than 500 stable cell lines, of which more than 100 can be used for antibody drug development, such as PD-1, PD-L1, Tim3, CTLA4, Lag3, Siglec15 as well as multiple GPCR cell lines.

Part of the stable cell lines developed and available for antibody drug development

  • Type Cell Line
    Antigen CHO-K1/siglec 15
    Antigen CHO-K1/mouse siglec 15
    Antigen CHO-K1/cyno siglec 15
    Antigen CHO-K1/CD73
    Antigen CHO-K1/Mouse CD73
    Antigen CHO-K1/Cyno CD73
    Antigen CHO-K1/NKG2D
    Antigen CHO-K1/cyno NKG2D
    Antigen CHO-K1/LAG3
    Antigen CHO-K1/Claudin18.2
    Antigen HEK293/Claudin18.2
    Antigen BxPc-3/Claudin18.2
    Antigen HEK293/mouse Claudin18.2
    Antigen HEK293/Claudin18.1
    Antigen CHO-K1/cyno CD40
    Antigen CHO-K1/cyno CXCR4
    Antigen CHO-K1/CD70
  • Type Cell Line
    Immune Checkpoint CHO-K1/PD-1
    Immune Checkpoint CHO-K1/mouse PD-1
    Immune Checkpoint CHO-K1/cyno PD-1
    Immune Checkpoint Daudi/PD-1
    Immune Checkpoint CHO-K1/PD-L1
    Immune Checkpoint CHO-K1/mouse PD-L1
    Immune Checkpoint CHO-K1/cyno PD-L1
    Immune Checkpoint Raji/PD-L1
    Immune Checkpoint CHO-K1/CD47
    Immune Checkpoint CHO-K1/Mouse CD47
    Immune Checkpoint CHO-K1/Cyno CD47
    Immune Checkpoint CHO-K1/CTLA4
    Immune Checkpoint CHO-K1/mouse CTLA4
    Immune Checkpoint CHO-K1/cyno CTLA4
    Immune Checkpoint CHO-K1/4-1BB
    Immune Checkpoint CHO-K1/mouse 4-1BB
    Immune Checkpoint CHO-K1/cyno 4-1BB
Service Highlights

1.One-stop service: Service package including gene synthesis, vector construction, lentivirus packaging, and host cell characterization to ensure the delivery of cell lines with stable expression of genes inserted. All we need is your gene name of interest.

2.Customized construction of expression vectors: Multiple promoters or co-expression of two or more genes in one plasmid. Various promoters and tags are available for your choice.

3.Multiple choices of expression validation methods: qPCR, RT-PCR, ELISA, Immunofluorescence, FACS and Western Blot.

4.Quality check: Cell line delivered is fungus, bacteria and mycoplasma free, and percentage cell viability is more than 90%.

5.Quality assurance: Valid source of the cell lines, traceable and complete experiment records.

6.Short turnaround time: Single cell clones can be delivered in just as short as three months.

Case study 1: Expression of unstable proteins

A common problem when developing stable cell lines for overexpression of gene of interest, which protein expression level are relatively low mRNA level are high. Rich experience has been accumulated in our trial and error to optimize the gene regulation and post-translation modification, enabling us stabilize the protein expression in need.

Case study 2: Expression of secreted proteins on cell membrane

Anchoring secreted proteins on the cell surface is a common way to accelerate the antibody drug screening. One example is to establish a stable cell line for tumor necrosis factor-α (TNF-α) that is mainly secreted by macrophages and responsible for inflammation.

Case study 3: Discriminative isoform-specific antibody development

Targeting protein isoform is challenging in antibody development given its difficulty to avoid off-target. By generating stable cells with protein isoforms respectively, antibody development for isoform-specificity can be neatly validated using FACS binding assay against target overexpression cell lines. One successful case is anti-Claudin 18.2 development whose isoform Claudin 18.1 is ubiquitously expressed in normal tissue.

The Stable Cell Line Service

Milestone Service Detail TAT (weeks)*
1 Host cell characterization Testing the single cell growth, killing curve 3~4
2 Expression vector construction Codon optimization, promoter selection, gene synthesis, plasmid preparation
3 Lentivirus generation Lentivirus packaging (optional) 1~2
4 Stable pool generation Cell pool with selection marker 3
5 Single cell clone generation and Screening Limited dilution to generate a single clone 3~4
6 Clone expansion, cryopreservation and QC Expanding the clone and preparing cryopreservation Check for bacteria, fungi and mycoplasma contamination 2~3
total 11~15

*The timeline is for estimation only, the actual delivery time may vary due to the complexity of biological process.