Evidence map›Paper›PMID 40407690›Full record

ArticleAntibodies (Basel, Switzerland)2025

A Targeted Integration-Based CHO Cell Platform for Simultaneous Antibody Display and Secretion.

Jessica P Z Ng, Mariati Mariati, Jiawu Bi, Matthew Wook Chang, Yuansheng Yang

Abstract read
In one paragraph

Article in Antibodies (Basel, Switzerland), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

What it found

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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

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3 · Its place in the literature

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0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Jessica P Z NgBioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #03-01 Centros, Singapore 138668, Singapore.
Mariati MariatiBioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #03-01 Centros, Singapore 138668, Singapore.
Jiawu BiInstitute of Molecular Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Dr, #07-01 Proteos, Singapore 138673, Singapore.ORCID 0000-0002-3735-0640
Matthew Wook ChangSynthetic Biology Translation Research Programme, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117465, Singapore.
Yuansheng YangBioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (A*STAR), 20 Biopolis Way, #03-01 Centros, Singapore 138668, Singapore.ORCID 0000-0002-0026-7069

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

objectiveWe developed a targeted integration-based CHO cell platform for simultaneous antibody display and secretion, enabling a streamlined transition from antibody library screening to production without requiring the re-cloning of antibody genes.

methodsThe platform consists of a CHO master cell line with a single-copy landing pad, a helper vector expressing FLPe recombinase, and bi-functional targeting vectors. Recombinase-mediated cassette exchange was utilized to integrate targeting vectors into the landing pad. Bi-functional vectors were designed by incorporating a minimal furin cleavage sequence (mFCS), RRKR, and various 2A peptides between the heavy chain (HC) and a membrane anchor.

resultsIncomplete cleavage at the mFCS and 2A sites facilitated the expression of both membrane-bound and secreted antibodies, while mutations in the 2A peptide produced a range of display-to-secretion ratios. However, a fraction of secreted antibodies retained 2A residues attached to the HC polypeptides. Further analysis demonstrated that modifying the first five amino acids of the 2A peptide significantly influenced furin cleavage efficiency, resulting in different display-to-secretion ratios for targeting vectors containing mFCS-2A variant combinations. To overcome this, we designed nine-amino-acid FCS variants that, when placed between the HC and membrane anchor, provided a range of display-to-secretion ratios and eliminated the issue of attached 2A residues in the secreted antibodies. Vectors with lower display levels proved more effective at distinguishing cells expressing high-affinity antibodies with closely matched binding affinities. The platform also demonstrated high sensitivity in isolating high-affinity antibody-expressing cells and supported robust antibody production.

conclusionThis targeted integration-based CHO platform enables efficient, in-format screening and production of antibodies with tunable display-to-secretion profiles. It provides a powerful and scalable tool for accelerating the development of functional, manufacturable therapeutic antibodies.

Indexed as

antibodiesCHO cellsfunctionalitymanufacturabilityrecombinase-mediated cassette exchangesimultaneous display and secretion

Identifiers

PMID40407690
PMCPMC12101391

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.