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

Gebhardt, Lauren; Abel, Molica; Zhou, Jing; Vogt, Audrey M.; Shin, Bo Hee; Herrick Wagman, Sarah L.; Santos, Ana; Puginier, Jerome; Wurm, Florian M.; Wurm, Maria J.; Yan, Guoying Grace; Adeniyi, Adedolapo; Lim, Sean K. H.; Somers, Will; Lin, Laura; D’Antona, Aaron M.; Zhong, Xiaotian

doi:10.3390/antib14040087

Open AccessEditor’s ChoiceArticle

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

by

Lauren Gebhardt

^1,†,

Molica Abel

^1,†,

Jing Zhou

¹,

Audrey M. Vogt

¹,

Bo Hee Shin

¹,

Sarah L. Herrick Wagman

¹,

Ana Santos

²,

Jerome Puginier

²,

Florian M. Wurm

^2,3

,

Maria J. Wurm

^2,4,*,

Guoying Grace Yan

¹,

Adedolapo Adeniyi

¹,

Sean K. H. Lim

¹,

Will Somers

¹,

Laura Lin

¹

,

Aaron M. D’Antona

¹

and

Xiaotian Zhong

^1,*

¹

BioMedicine Design, Preclinical and Translational Sciences, Pfizer Research and Development, 610 MainStreet, Cambridge, MA 02139, USA

²

MagellanBiologics, Parque Tecnológico de Cantanhede Edificio III Laboratório 5, 3060-197 Cantanhede, Portugal

³

Life Science Faculty, Swiss Federal Institute of Technology Lausanne, 1015 Lausanne, Switzerland

⁴

ExcellGene SA, Route d’ile au Bois 1a, 1870 Monthey, Switzerland

^*

Authors to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Antibodies 2025, 14(4), 87; https://doi.org/10.3390/antib14040087

Submission received: 5 August 2025 / Revised: 17 September 2025 / Accepted: 9 October 2025 / Published: 11 October 2025

(This article belongs to the Special Issue A Festschrift Celebrating Dr. Dimiter Stanchev Dimitrov: Antibodies, Innovation, and Impact on Infectious Disease and Cancer Research)

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Abstract

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 Pure^TM 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.

Keywords: transient gene expression; in-culture antibody capturing; CHO4Tx^®; magnetic ProA beads; semi-automation; GenScript AmMag™ SA-Plus; AKTA Pure^TM

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MDPI and ACS Style

Gebhardt, L.; Abel, M.; Zhou, J.; Vogt, A.M.; Shin, B.H.; Herrick Wagman, S.L.; Santos, A.; Puginier, J.; Wurm, F.M.; Wurm, M.J.; et al. A Robust, High-Titer, Semi-Automated, and In-Culture Antibody-Capturing Transient CHO Platform Technology. Antibodies 2025, 14, 87. https://doi.org/10.3390/antib14040087

AMA Style

Gebhardt L, Abel M, Zhou J, Vogt AM, Shin BH, Herrick Wagman SL, Santos A, Puginier J, Wurm FM, Wurm MJ, et al. A Robust, High-Titer, Semi-Automated, and In-Culture Antibody-Capturing Transient CHO Platform Technology. Antibodies. 2025; 14(4):87. https://doi.org/10.3390/antib14040087

Chicago/Turabian Style

Gebhardt, Lauren, Molica Abel, Jing Zhou, Audrey M. Vogt, Bo Hee Shin, Sarah L. Herrick Wagman, Ana Santos, Jerome Puginier, Florian M. Wurm, Maria J. Wurm, and et al. 2025. "A Robust, High-Titer, Semi-Automated, and In-Culture Antibody-Capturing Transient CHO Platform Technology" Antibodies 14, no. 4: 87. https://doi.org/10.3390/antib14040087

APA Style

Gebhardt, L., Abel, M., Zhou, J., Vogt, A. M., Shin, B. H., Herrick Wagman, S. L., Santos, A., Puginier, J., Wurm, F. M., Wurm, M. J., Yan, G. G., Adeniyi, A., Lim, S. K. H., Somers, W., Lin, L., D’Antona, A. M., & Zhong, X. (2025). A Robust, High-Titer, Semi-Automated, and In-Culture Antibody-Capturing Transient CHO Platform Technology. Antibodies, 14(4), 87. https://doi.org/10.3390/antib14040087

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

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