Stable Cell Line Services

Background

Overview of Service

Stable Cell Line Development for Precision Research

Creative BioMart delivers rigorously optimized stable cell line generation services to accelerate your protein therapeutics development, functional genomics studies, and CRISPR-based applications. Our scientifically validated workflows integrate proprietary expression vectors, advanced selection markers (e.g., antibiotic/fluorescence-based), and high-throughput screening technologies to produce clonal cell lines with consistent protein expression, targeted gene knockdown, or precise genome editing outcomes.

Unlock Your Research Potential Today

From exploratory gene editing projects to GMP-ready cell line development, Creative BioMart’s end-to-end solutions balance scientific rigor with budget-conscious pricing. Contact our cell biology specialists to design a workflow aligned with your therapeutic discovery, biosimilar development, or functional genomics objectives.

Service Highlights

• Precision Vector Engineering & Tailored Genetic Constructs

  Creative BioMart delivers precision-engineered vector design solutions integrating codon optimization algorithms, promoter selection (CMV, EF1α, inducible systems), and genomic stability elements to maximize transgene expression and clonal integrity. Our proprietary expression vectors are validated across HEK293, CHO, and custom host systems, supporting diverse applications from therapeutic antibody production to CRISPR-mediated gene editing. For specialized needs, we provide customized solutions to you.

• Accelerated Clone Screening & Scalable Compliance-Ready Platforms

  Accelerate timelines with our robotics-enabled high-throughput screening platform, which identifies high-expressing clones via fluorescence-activated sorting (FACS) and mini-batch expression profiling. All workflows adhere to animal-component-free standards, eliminating viral/bovine contamination risks while ensuring regulatory compliance. Backed by 150+ successfully developed cell lines, our end-to-end support spans construct design, clone validation (qPCR, Western blot), and master cell bank establishment.

Service Details

1

Project Initiation

Customers submit target gene sequence or protein information to confirm cell line selection and service needs.

Timeline: 1-2 weeks

2

Vector Construction

Gene optimization, vector construction and sequence verification.

Deliverables:

Sequencing results

Timeline: 1-2 week

3

Cell Transfection

Lentivirus packaging, virus transduction, plasmid transfection

Deliverables:

Transfection efficiency and cell growth status evaluation

Timeline: 2-3 weeks

4

Selection & Expansion

Antibiotic screening, drug screening and high throughput screening

Timeline: 6-8 weeks

5

Quality Validation

RT-qPCR, Mycoplasma testing, FACS or WB, Immunofluorescence

Deliverables:

Stability test result

Timeline: 1-2 weeks

6

Delivery & Support

Detailed project report, Follow-up technical support

Deliverables:

Frozen stable cell lines (at least 2 frozen tubes)

Timeline: 1 week

Core Services

Types	Details
Gene Overexpression Stable Cell Lines	Constitutive or inducible expression of single/multiple genes.
Gene Knockdown Stable Cell Lines	Using shRNA or CRISPRi for targeted gene suppression.
Gene Knock-out/Knock-in Stable Cell Lines	Precision gene editing with CRISPR/Cas9.
miRNA Overexpression Stable Cell Lines	For studying miRNA function.
Customizable Expression Systems	Including inducible systems (e.g., Tet-On/Tet-Off).
More Cell Lines	Protein Production Grade Stable Cell Lines; Assay Grade Stable Cell Lines

Service Delivery & Submission Guidelines

• What We Need From You

• Clean & Certified Cells
  • Share mycoplasma-free cells (tested within 3 months) – think of it as a “health certificate” for your project. If using third-party lines (e.g., HEK293, CHO), just forward their origin docs – we’ll handle the rest!
  • No cells yet? Let’s discuss! We source ATCC-certified lines or engineer custom hosts.
• DNA or Cells? Ship Like a Pro
  • DNA Route: 50 μL of gDNA (≥25 ng/μL) + buffer details → pack with ice, and we’ll optimize codon usage.
  • Live Cell Path: Ship ≥5 million viable cells (90%+ vitality) in their favorite medium – we’ll amplify them under your specified conditions (FBS%, CO₂ levels, etc.).
• Vector Compatibility Check
  • Using your plasmid? Share sequencing files – we’ll confirm resistance marker compatibility to avoid surprises.

• What You Get

• Cell Lines That Work Harder
  • Monoclonal Superstars: FACS-sorted single-cell clones with proof of monoclonality – because reproducibility starts with purity.
  • Ready-to-Express Pools: Need protein fast? Get polyclonal lines in 12 weeks, perfect for CRISPR knockout validation or rapid antibody production.
• Reports That Impress You
  • The Full Story: From vector maps to Western blot data, our CoA includes sterility checks and endotoxin levels (<0.1 EU/μg).
  • Validation Your Way: Pick qPCR, FACS, or ELISA – we’ll show your gene is expressed, stable, and ready for scale-up.
• Speed Without Compromise
  • Stable Pools: Protein in hand by Week 12.
  • Monoclonals: Clonal lines delivered by Week 16 (or sooner – ask about our priority track!).

Ready to Start? Let’ stailor a plan for your next therapeutic candidate, biosimilar, or functional genomics breakthrough.

Add-On Services

• Long-Term Stability Testing : 15-20 passage analysis for consistent expression.
• Scale-Up Support : Master cell bank development under GLP/GMP standards.
• Vector Design Assistance : Codon optimization, promoter engineering, or episomal system integration.
• CHO Stable Cell Line Development for Antibody/Protein Production : From codon-optimized vector design to high-titer monoclonal clones—for robust, scalable production of therapeutic antibodies, bispecifics, or complex glycoproteins. Click here to learn more about Mammalian Expression Systems .

Why Choose Us?

Superior Stability Superior Stability. After 25 generations, the protein expression level was still similar to that of the early generations.

High Protein Yield. Efficient expression of target proteins to meet production needs.

Fast Turnaround. Development of highly productive stable cell clones within 12 weeks.

Customized Solutions. Cell line development tailored to customer needs.

Rigorous QC. From gene verification to protein expression, quality is strictly controlled throughout.

Broad Applications. It is suitable for protein production, gene research, drug screening and other fields.

Case Study

* NOTE: We prioritize confidentiality in our services to safeguard technology and intellectual property for enhanced future value and protection. The following case study has been shared with the client's consent.

Project Name: Construction of Stable Cell Line for Antibody Expression
Goal
The project focuses on the construction of a stable cell line for the expression of a specific antibody. The antibody sequence information was provided by the customer. The primary objective is to develop a stable cell line that can efficiently express the target antibody.
Results

1. Protein Sequence : The amino acid sequences of the heavy chain and light chain were provided by the customer.
2. Monoclonal Stable Cell Line Screening : After transfecting the constructed plasmids into CHO cells and culturing for 6 days, the supernatant was collected and analyzed by Western Blot (WB). Two high-expressing CHO stable cell lines ( 9F6 and 9B1 ) were identified.

   

   Fig. 1. SDS-PAGE analysis of antibody expression.

   Lane M: Protein Marker; Lane 1: CHO cell 7B10 reduced; Lane 2: CHO cell 7B10 non-reduced; Lane 3: CHO cell 9F10 reduced; Lane 4: CHO cell 9F10 non-reduced; Lane 5: CHO cell 9F4 reduced; Lane 6: CHO cell 9F4 non-reduced; Lane 7: CHO cell 9E6 reduced; Lane 8: CHO cell 9E6 non-reduced; Lane 9: CHO cell 9E9 reduced; Lane 10: CHO cell 9E9 non-reduced; Lane 11: CHO cell 9B1 reduced; Lane 12: CHO cell 9B1 non-reduced; Lane 13: CHO cell 10F5 non-reduced; Lane 14: CHO cell 10F5 reduced; Lane 15: CHO cell 9F6 non-reduced; Lane 16: CHO cell 9F6 reduced; Lane 17: CHO cell 8E10 reduced; Lane 18: CHO cell 8E10 non-reduced; Lane 19: CHO cell 10F7 reduced; Lane 20: CHO cell 10F7 non-reduced; Lane 21: CHO cell 7B7 reduced; Lane 22: CHO cell 7B7 non-reduced.

   

   Fig. 2. SDS-PAGE analysis of antibody expression.

   Lane M: Protein Marker; Lane 1: CHO cell 9F6 reduced; Lane 2: CHO cell 9F6 non-reduced.

   

   Fig. 3. SDS-PAGE analysis of antibody expression.

   Lane M: Protein Marker; Lane 1: CHO cell 9B1 non-reduced; Lane 2: CHO cell 9B1 reduced.

3. Antibody Expression and Purification : The CHO cell 9F6 was selected for further purification due to its higher expression level. Purification was performed using a Protein A affinity column. The purified antibody was analyzed by SDS-PAGE and Western blot, confirming successful purification.

   

   Fig. 4. SDS-PAGE analysis of antibody purification.

   Lane M: Protein Marker; Lane 1: Purified atibody reduced; Lane 2: Purified atibody non-reduced.

   

   Fig. 5. Western blot analysis of antibody purification.

   Lane M: Protein Marker; Lane 1: Purified antibody non-reduced.

Conclusions

The antibody recombinant plasmids were successfully constructed and transformed into CHO-K1 cells. Two high-expressing stable cell lines were obtained, with CHO cell 9F6 showing higher expression levels. The purified antibody was measured by BCA assay, yielding 15.8 mg of antibody at a concentration of 1.58 mg/mL from 25 mL cell culture. The actual yield of CHO cell 9F6 was significantly higher than the expected 0.632 g/L, despite losses during purification. The goal of obtaining a stable cell line expressing the antibody was successfully achieved.

Customer Testimonials

"Creative BioMart’s CHO stable cell line development transformed our mAb project with optimized glycosylation (G0F >85%). Their monoclonal screening workflow and host cell protein (HCP) clearance strategies were pivotal in securing Phase I trial approvals."

-Dr. Sarah Johnson, Head of Biotherapeutics | Biopharma Innovator

"With their high-density fermentation expertise, the protein expression stability across 30+ passages ensured consistent enzyme activity (≥95%), accelerating our green chemistry initiatives."

-Mr. Carlos Rodriguez, CTO | Industrial Enzyme Producer

"Rapid stable cell pool development enabled same-week shipment of COVID-19 antigen reagents during peak demand. Their gene knockout validation (FACS/qPCR) met FDA EUA documentation standards flawlessly."

-Dr. Mei Ling, R&D Director | Diagnostic Reagent Company

"Custom CRISPR-edited HEK293 cell lines expressing ATP-binding cassette transporters exceeded our cryo-EM resolution needs. Creative BioMart’s stable cell line characterization (solubility >90%, confirmed activity via ATPase assays) provided publication-grade validation for our Cell manuscript."

-Prof. Alessandro Russo, PI | Academic Structural Biology Lab

"Outsourcing GMP-compliant cell line development cut costs by 35% versus in-house efforts. Their master cell banks passed EMA audits, and their CHO cell line platform supported bispecific antibody projects."

-Ms. Fatima Al-Mansoori, VP Operations | CDMO specializing in Biosimilars

"Creative BioMart’s secretory expression system in plant suspension cells achieved 80% soluble yields for our eco-friendly protein. Their custom solutions bridged academic proof-of-concept to pilot-scale production."

-Dr. Ethan Cole, CSO | AgriTech Biomanufacturing Firm

FAQs

• Q: What types of stable cell lines do you develop for protein production?

  A: We engineer custom stable cell lines across multiple platforms to match your project needs:

  • CHO & HEK293 systems: For high-yield therapeutic antibodies, cytokines, or complex glycoproteins with human-like post-translational modifications.
  • Insect cell lines (Sf9/Sf21): Baculovirus -driven expression of multi-subunit proteins or viral-like particles.
  • Specialized hosts: Suspension-adapted lines for scalable production or engineered lines for secretory/membrane protein challenges.

• Q: How do you ensure stable, high-yield protein expression in engineered cell lines?

  A: Our proprietary workflow guarantees reliability:

  • Vector optimization: Codon usage tailored to your host, promoter selection (CMV, EF1α), and integration methods (CRISPR) for genomic stability.
  • AI-driven clone screening: Machine learning prioritizes high producers using early-stage viability and productivity markers.
  • Multi-passage validation: Expression consistency verified over 20+ generations via qPCR, Western blot, and activity assays.

• Q: Can you handle complex proteins prone to aggregation or insolubility?

  A: Every cell line undergoes:

  • Purity testing: Mycoplasma-free certification, endotoxin levels (<0.1 EU/μg), and sterility checks.
  • Functional validation: Target-specific assays (e.g., ELISA, SPR for binding affinity, glycan profiling via HPLC).

• Q: Can you support niche projects like toxic protein production or inducible systems?

  A: Yes. Our custom solutions tackle complex challenges:

  • Toxicity mitigation: Inducible promoters (Tet-On/Off), stress-response engineering, or fed-batch media optimization.
  • CRISPR-edited lines: Knock-in fluorescent tags, secretion signals, or chaperone co-expression for difficult targets.
  • Viral vector packaging: HEK293-derived lines for high-titer AAV/lentivirus production with optimized transfection efficiency.

• Q: What timelines can we expect for stable cell line development?

  A: Rapid cell pools: 10-12 weeks for initial protein supply (e.g., research-scale antibodies).
  Monoclonal lines: 16-18 weeks with full QC, extendable for glycoengineering or inducible systems.
  Priority projects: Accelerated timelines available for urgent IND/CTA-enabling studies.