● 4-chain BsAb: 8.3 g/L (with 88% correct pairing)
● ADC mAb: 10 g/L
● IND-Filed Molecule: 15 g/L at 500L scale
● Nanobody: 13.1 g/L
● Probody: 8.0 g/L
● Hard Expressed Proteins Fusion Proteins: 4.2 g/L
Fluorescent Site-Specific Integration
Cell Line Development
The site-specific integration cell line development platform, AlfaCell, integrates fluorescence-based visual selection with site-specific integration (SSI) technology to enable precise insertion of genes of interest into stable, high-titer, and high-quality genomic hot spots. The platform enables rapid generation of high-yield and stable cell lines within 1.5 months, and early prediction of product quality within 1 month, empowering clients to accelerate the development of cell lines ready for large-scale industrial manufacturing.
Technical Background
Recombinant Therapeutic Proteins (RTPs) require correct folding and post-translational modifications for activity. While traditional random integration is simple, it often results in low yield, long timelines, high costs, and unpredictability. SSI technology addresses these issues by enabling targeted gene insertion into validated active loci via homologous recombination, advancing precise and efficient cell line development.
Site-specific integration (SSI) is a recombinase-mediated genome editing method that inserts exogenous genes into specific, predetermined genomic sites of host cells in a targeted manner. It achieves controllable single-copy gene insertion, effectively overcoming the positional variation and expression instability of randomly integrated cell clones, and has become a core tool for high-stability, regulatory-friendly mammalian cell line development for therapeutic protein manufacturing.
Services
We perform services in our facility
Using the AlfaCell platform, we develop a cGMP cell line specifically for your program and ship it to you.
We transfer AlfaCell platform to your facility
We ship you our AlfaCell host cell and you use our platform to perform cGMP cell line development in your lab.
| Pain Points | Site-specific Integration Platform | Transposon System |
Random Insertion (CHO-K1 WT and CHO-GS) |
|---|---|---|---|
| Titer | 4–16 g/L | 3–5 g/L | 2–5 g/L |
| Timeline |
1.5 months (due to green protein application) |
> 6 months | > 6 months |
| Stability | Pre-selected single stable site | Unpredictable due to multi-sites | Unpredictable due to random insertion |
| Workload | Screened with 1–2 of 96-well plates | Screened with 50–100 of 96-well plates | Screened with 50–100 of 96-well plates |
| Product Quality Evaluation |
Able to evaluate product qualities from homogenized cell pool within 1 month post transfection |
Unable to evaluate product qualities until RCB available (within 3–4 months) |
Unable to evaluate product qualities until RCB available (within 3–4 months) |
Advantages
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Case Study
High-Titer
High Quality
High Speed
Powered by site-specific integration and AI intelligent positioning technologies, the AlfaCell platform precisely inserts target genes into validated high-expression genomic hotspots. It supports the generation of high-yield and stable cell lines within 1.5 months, avoiding the instability and repetitive screening work of conventional random integration strategies. Meanwhile, product quality can be preliminarily assessed in the first month, delivering industrial-grade monoclonal cell lines ready for scale-up production to global biotech partners。
Stability
The clone stability acceptable with same integrated site.
Evaluate clone stability.
Easy Transfer
The AlfaCell platform enables seamless, reliable tech transfer from GBB Lab to factories and CDMO partners. Engineered for easy scale-up, our processes ensure consistent high-yield replication with minimal adaptation, completing transfer in just ~28 days. This streamlined pathway accelerates your commercial manufacturing timeline while mitigating scale-up risks.
Applications
Monoclonal Antibody
Stable integration ensures consistency in key quality attributes (CQAs), including charge heterogeneity and glycosylation modifications, across production batches.
TCE Drug
The AlfaCell platform completed the cell line development for this TCE drug, achieving a titer of 5.6 g/L and a correct pairing rate of 84%.
Four-chain BsAb
In just 3 months, GBB successfully completed the cell line development for this four-chain BsAb, achieving a titer of 8.3g/L and a correct paring rate of 87.9%.
Nano Bispecific Antibody
Within just 3 months, GBB completed stable cell line development for Co. B's nano bispecific antibody. The top-performing cell line achieved a titer of 15.6 g/L, successfully meeting and exceeding the industrial production target.
Site-Specific Integration Unlocks Step-Change
After adopting AlfaCell’s site-specific integration technology, the bispecific antibody achieved a titer of 4 g/L within just 1.5 months, with a favorable growth profile.
Parallel Cell Line Development Prior to Final PCC Confirmation
Based on the results of AlfaCell transient expression (molecules + optimized plasmids), the AlfaCell platform constructs and screens stable cell lines for multiple molecules in parallel. The platform’s unique "Double-Fast" advantage enables obtaining high-titer and stable cell lines within 1.5 months and rapid prediction of product quality within 1 month, allowing early planning for subsequent process development and capacity expansion.
