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Still using cell-based assays for screening? This lab has already leapfrogged into antibody development with a “cell-free” approach.
Release date:
2026-04-14
Traditional antibody discovery resembles a marathon: construction, transfection, expression, purification, and screening—processes that can span weeks or even months, with limited throughput and low success rates for difficult-to-express antibodies or toxic proteins. Moreover, many potentially highly active antibodies are mistakenly classified as “negative” due to insufficient expression levels, resulting in the loss of promising candidates. In contrast, cell-free protein synthesis (CFPS) technology creates an “ultra‑fast pathway” for antibody screening, compressing workflows that traditionally take weeks or months into just days. Drawing on existing literature, CFPS has demonstrated two key advantages in antibody screening:
1. High yield and high activity coexist
Gang et al. employed an E. coli–based cell-free synthesis system (OCFS, a type of CFPS technology) to successfully produce correctly folded, fully functional, non‑glycosylated full-length trastuzumab, which exhibited antigen‑binding affinity comparable to that of antibodies produced in mammalian cells. These antibodies can be rapidly scaled up in standard bioreactors, with linear expansion achievable within hours and yields reaching up to hundreds of milligrams per liter—representing a more than one‑million‑fold range of scale—thus demonstrating their utility as a seamless platform spanning antibody discovery through preclinical development [1].
Figure 1: Schematic Diagram of the OCFS System
2. Ultra-fast filtering, doubling efficiency
An integrated platform based on CFPS enables end-to-end automation, from DNA template assembly and protein synthesis to binding‑activity assays. In tests targeting the SARS‑CoV‑2 spike protein, Hunt et al. completed the expression and functional evaluation of hundreds of antibodies in under 24 hours, successfully identifying all neutralizing antibodies that have received Emergency Use Authorization (EUA) and discovering novel candidate molecules. Compared with the weeks required by conventional methods, this approach achieves an efficiency improvement of several orders of magnitude [2].
Figure 2: Workflow for Rapid Cell-Free Antibody Screening
Ojima, Kato, and colleagues have developed a high-throughput, low-cost monoclonal antibody screening method—“Ecobody”—which integrates single B-cell sorting with cell-free protein synthesis (CFPS) technology. This approach enables the entire workflow, from isolating B cells from immunized animals to screening and characterizing antigen-binding fragments (Fab), to be completed within two working days, representing an order-of-magnitude improvement in speed compared with conventional methods [3].
Figure 3: Ecobody technology for the rapid screening of monoclonal antibodies from single B cells using CFPS.
Success Stories
PLD Technology has successfully established a high-throughput cell-free protein expression platform capable of screening 3,000 antibodies per day, with fully automated synthesis and no manual intervention. Antibody screening is now no longer limited to weekly or even monthly cycles. Our proprietary CFPS system supports the autonomous assembly of multiple polypeptide chains, enabling soluble expression in the supernatant of antibody fragments such as VHHs and scFvs, as well as full-length IgG antibodies. ELISA assays demonstrate that the antibodies produced by our company exhibit high immunological activity.
PLD High-Throughput Cell-Free Protein Expression Workstation
Figure: SDS-PAGE analysis of an IgG and its Fab fragment, along with their ELISA activity (CFPS)
References:
- Yin, Gang et al. “Aglycosylated antibodies and antibody fragments produced in a scalable in vitro transcription-translation system.” mAbs vol. 4,2 (2012): 217-25. doi:10.4161/mabs.4.2.19202
- Hunt, Andrew C et al. “A rapid cell-free expression and screening platform for antibody discovery.” Nature communications vol. 14,1 3897. 3 Jul. 2023, doi:10.1038/s41467-023-38965-w.
- Ojima-Kato, Teruyo et al. “Ecobody technology: rapid monoclonal antibody screening method from single B cells using cell-free protein synthesis for antigen-binding fragment formation.” Scientific reports vol. 7,1 13979. 25 Oct. 2017, doi:10.1038/s41598-017-14277-0.
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