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  4. A Robust, High-Titer, Semi-Automated, and In-Culture Antibody-Capturing Transient CHO Platform Technology
 
research article

A Robust, High-Titer, Semi-Automated, and In-Culture Antibody-Capturing Transient CHO Platform Technology

Gebhardt, L. P.
•
Abel, Molica
•
Zhou, Jing
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October 11, 2025
Antibodies

Background: Recent advances in antibody discovery technologies, especially progress in de novo synthesis through machine learning, have imposed a significant production challenge for the generation of a large diversity of antibodies against nearly any target of interest. There is a demand for the rapid production of dozens of purified antibodies in 10-milligram quantities sufficient for functional screening and molecular assessment studies. Objectives: To meet this requirement, a semi-automated production methodology and workflow was developed to bridge the miniaturized high-throughput screenings (HTSs) and the conventional custom-scale workflow by taking advantage of four new technology applications. Methods: First, it exploited a novel, simple, high-titer transient expression system, “CHO4Tx®”, which could achieve high yields in the range of 200 mg/L and above, across a variety of antibody constructs, including challenging targets. The consistently high yields from this transient CHO platform enabled the delivery of ~20 mg of crude material per 100 mL scale flask production with a throughput capacity of nineteen constructs in a single run. Secondly, we established a magnetic ProA bead in-culture antibody-capturing process, which significantly shortened the production timeline by eliminating the steps of cell centrifugation, filtration, and medium column loading. Third, we utilized the GenScript AmMag™ SA Plus semi-automation, which could handle magnetic ProA bead elution for 12 constructs within less than 1 h. Lastly, we transformed the AKTA PureTM system into an automated buffer exchange purification system with a capacity of processing 19 samples in a single run. Results and Conclusions: This new production platform was proven to be robust and could be applied for the routine production of antibodies of sufficient quality and quantity in support of cell-based assays and biophysical characterization.

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Type
research article
DOI
10.3390/antib14040087
Author(s)
Gebhardt, L. P.

Pfizer (United States)

Abel, Molica

Pfizer (United States)

Zhou, Jing

Pfizer (United States)

Vogt, A

Pfizer (United States)

Shin, Bo Hee

Pfizer (United States)

Wagman, Sarah L. Herrick

Pfizer (United States)

Santos, Ana Luísa
Puginier, Jerome
Wurm, Florian Μ.  

École Polytechnique Fédérale de Lausanne

Wurm, Florian Μ.  

École Polytechnique Fédérale de Lausanne

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Date Issued

2025-10-11

Publisher

MDPI AG

Published in
Antibodies
Volume

14

Issue

4

Start page

87

Subjects

transient gene expression

•

in-culture antibody capturing

•

CHO4Tx ®

•

magnetic ProA beads

•

semi-automation

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GenScript AmMag™ SA-Plus

•

AKTA Pure TM

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
EPFL  
Available on Infoscience
October 20, 2025
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/255058
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