Are you struggling with long timelines and inconsistent results in your monoclonal antibody development? Leveraging our revolutionary Verified In-Situ Plate Seeding (VIPS™) technology, we guarantee efficient single-cell seeding with image-based proof of clonality, slashing development time by over 50% while providing clonality reports for successful IND submission. Opt for our proprietary, IP-free cell lines, designed for stable development and provided royalty-free. We tailor each project to your specific needs, from monoclonal isolation to comprehensive productivity assessments, and deliver a detailed monoclonality report that surpasses industry standards. Take your research to production efficiently and confidently with ProteoGenix’s proven expertise.

Comprehensive Custom Cell Line Development workflow

Expression vector construction

  • Gene synthesis with codon optimization for the target expression system.
  • High-performance expression vector subcloning.

Developability Study

  • Production testing in transient expression systems.
  • Biochemical characterization properties and full reporting.

Host Cell Transfection and Characterization of Stable Transfected Pools

  • Host cell stable transfection with recombinant plasmid.
  • Generation and characterization of stable transfected pools.
  • Selection and amplification of the most productive stable pools.

Best Monoclones Isolation and Selection by VIPS™ Technology

  • Monoclone isolation using Verified In-Situ Plate Seeding (VIPS™).
  • Screening and selection of highest producing clones via ELISA and other assays.

Best Monoclones Characterization

  • Productivity evaluation in batch and/or fed-batch cultures.

RCB QC and Stability Study

  • QC analysis and custom cell line stability study.
  • Transfer of RCB and optimized protocols.
Step Content Timeline Deliverables
Gene Synthesis and Cloning
  • Synthesis of coding sequence(s)
  • Subcloning into expression vector
~1 week
  • Cloning plasmids
  • DNA sequences of coding regions
Transient Expression Evaluation
  • Plasmid amplification
  • Transfection of XtenCHO cells
  • Small scale expression and purification
~2 weeks
  • Production yield and biochimecal properties report
  • Purified sample (optional)
Generation of Stable Pools
  • Generation and selection of CHO cell pools
  • Antibody integrity assessments and quantification
  • RCB of the best stable pool
  • Genetic material production, cell transfection, cell culture and Ab titration to select polyclonal pool(s) report
  • Antibody production and characterization report
Single Cell Clone Screening by VIPS™
  • Monoclonal isolation using VIPS™
  • Monoclonality evaluation
  • Expansion and expression screening of top clones
~8-10 weeks
  • Monoclonality report for confident IND submission
  • Top clone comparison report
  • Purified antibodies from selected clonesl
RCB Preparation and QC
  • Culture and quality control of top clones
  • Freezing of Research Cell Banks (RCB)
~1 weeks
  • Certificate of analysis of RCBs
  • Development of custom cell line, selection, and RCB report
RCB Stability Study
  • Subculturing and productivity evaluation of clones every 10 generation
~ 4-5 weeks
  • Stability study report
  • Clone selection recommendation
  • Comprehensive antibody analytics: affinity, aggregates, thermostability tests
~ 2-3 weeks
  • Detailed analytics reports
  • Affinity and stability test results

Options:Further evaluation and characterization of the antibody can be done by

  • Analysis of Antibody Aggregates (SEC-HPLC)
  • Endotoxin detection test
  • IgG Quantification (UV280)
  • Activity analysis by ELISA against the antigen
  • DSC Analysis (Thermostability Analysis by Differential Scanning Calorimetry)
  • Affinity Determination (Kd) against soluble antigen via Biacore X100
  • Affinity Determination (Kd) against antigen expressed on cell surface via Surface Plasmon
  • Resonance imaging (SPRi)
  • FACS analysis
  • Biological potency (ADCC assay)
  • Glycosylation profile (LC/MS)

Customization and Flexibility in Cell Line Generation Service

ProteoGenix is committed to meeting the unique needs of each client by offering customizable services that go beyond standard offerings. Here’s how we tailor our services :

  • In Silico Antibody Developability Assessment: We conduct preliminary in silico studies to evaluate antibody developability, producing and characterizing variants to identify the most stable sequence for cell line development.
  • Detailed Milestones and Reporting: Clients are kept in full control of their projects through detailed milestones, comprehensive reporting, and strategic go/no-go decision points, allowing them to track progress and make informed decisions throughout the development process.
  • Complete CMC Package: We provide a comprehensive Chemistry, Manufacturing, and Controls (CMC) package for thorough characterization of the antibodies produced.

VIPS™ Technical Specifications and Advantages

The use of Verified In-Situ Plate Seeding (VIPS™) technology in custom cell line development offers several significant technical advantages:

  • High-Efficiency Cloning: VIPS™ technology enables high-efficiency single-cell seeding, crucial for generating monoclonal cell lines. This automated process significantly reduces the manual labor and potential for error associated with traditional methods like limiting dilution cloning, thus ensuring more reliable and faster cell line development.
  • Assured Clonality: VIPS™ provides image-based evidence of clonality right from day zero. This “double-lock” approach ensures that each cell line starts from a single, visually confirmed cell, meeting stringent regulatory requirements for monoclonality. This is critical not only for scientific validation but also for regulatory approvals in biopharmaceutical production.
  • Time and Cost Reduction: By automating the seeding and initial growth stages, VIPS™ cuts down the time required to establish stable cell lines. Traditional methods can be time-consuming and labor-intensive, but VIPS™ streamlines this process, leading to significant reductions in both time and operational costs associated with cell line development.
  • Enhanced Outgrowth and Stability: The precise delivery and verification of single-cell seeding improve the outgrowth and stability of the clones. VIPS™ ensures that the cells grow from a verified single-cell origin, which enhances the uniformity and reproducibility of the cell lines developed. This uniformity is crucial when these cell lines are scaled up for therapeutic protein production, as it ensures consistent product quality.
  • Regulatory Compliance: The detailed documentation and clonality reports generated by VIPS™ facilitate compliance with regulatory standards. These reports provide a reliable audit trail that can be critical during the review process by regulatory bodies, thus de-risking the submission and approval phases of biopharmaceutical development.

Custom Cell Line Development : Expression Systems and Formats

Proprietary CHO

  • Amplification and Selection: Available with both MTX/DHFR-mediated or Methionine Sulfoximine (MSX)/GS-mediated amplification and selection.
  • Advantages: Offers freedom to operate with a one-time fee payment, making it cost-effective for long-term use.
  • Applications: Suitable for the rapid and easy transfer for cGMP production, supporting scalable and flexible biomanufacturing processes.


  • Amplification and Selection: Utilizes MTX/DHFR-mediated or Methionine Sulfoximine (MSX)/GS-mediated amplification and selection.
  • Advantages: No licence fee is required before moving towards commercial use, reducing initial investment costs.
  • Applications: Designed for straightforward scale-up and transfer, facilitating seamless progression from research to commercial production.


  • Amplification and Selection: Employs MTX/DHFR-mediated amplification and selection.
  • Advantages: No license fees for early-stage use and is renowned for producing FDA-approved biotherapeutics.
  • Applications: Ideal for the bioproduction of therapeutic proteins under stringent regulatory requirements.

HEK293 Variants (HEK293, 293F, 293E)

  • Characteristics: These are human embryonic kidney cells that are versatile for transient and stable expression with high yield.
  • Advantages: HEK293 cells are known for rapid growth and high protein yield, making them suitable for both research and commercial scale productions.
  • Applications: Commonly used in the production of viral vectors for gene therapy, vaccines, and recombinant protein products.

Customer-Specific Cell Lines

  • Customization: We offer the flexibility to develop and utilize custom cell lines as per client-specific requirements.
  • Advantages: Tailored to meet unique project needs, ensuring optimal expression and productivity.
  • Applications: Custom cell lines are particularly useful for specialized therapeutic targets or when proprietary systems are needed for competitive differentiation.

Case Study

Gene Synthesis, Subcloning, And Transient Expression

Transient transfection in CHO-K1 cells (30 ml) and purification using protein G resin.

Yield: 18.2 mg/L
Quantity Produced: 0.18 mg
Purity: >90%

A final QC was performed by reduced and non-reduced PAGE analysis confirming the integrity of the antibody

Development Of Stable-Transfected Pools

Construction of stable expression vectors (one for the heavy chain and another for the light chain) followed by vector linearization and transfection.

Selection in the presence of MSX : 3

Pool ID Quantity (mg) Yield (mg/L) Purity
1 37.3 1243.0 >90%
2 39.4 1313.3 >90%
3 43.6 1453.3 >90%

Production And Purification Scale-Up

1L fed-batch production was carried out in 3L flasks. Purification was performed using protein A resin.

2015.5 mg/L >95% purity

Isolation And Screening Of Stable Monoclones

Monoclones were isolated using the limiting dilution method in 96-well plates and expression evaluation was performed with ELISA with anti-Fc antibodies.

2 Rounds of limiting dilution

29 Best performing clones

Best monoclones were evaluated by SDS-PAGE and ELISA (Fc-antibody). ELISA data (below) revealed 10 monoclones with high expression levels.

Case study Custom cell line development
Clone ID 1 3 4 7 10 15 17 19 30 31
Yield (g/L) 2.05 2.86 1.07 1.68 4.34 2.57 3.21 2.60 1.45 1.25

Monoclones 3, 10, 17,19 maintained good stability and viability in subsequent tests.

Scale-Up Production

Monoclonal 10 was used for scale-up and achieved excellent yields with optimized culture conditions.

7.91 g/L >95% purity

Stable Cell line development Service at ProteoGenix: FAQ