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Cross-Interaction Chromatography: A Rapid Method to Identify Highly Soluble Monoclonal Antibody Candidates

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Abstract

Purpose

To develop a high-throughput cross-interaction chromatography screening method to rapidly identify antibody candidates with poor solubility using microgram quantities of purified material.

Methods

A specific recombinant antibody or bulk polyclonal IgG purified from human serum was chemically coupled to an NHS-activated chromatography resin. The retention times of numerous monoclonal antibodies were determined on this resin using an HPLC and compared to the solubility of each antibody estimated by ultrafiltration.

Results

Retention times of the antibodies tested were found to be inversely related to solubility, with antibodies prone to precipitate at low concentrations in PBS being retained longer on the columns with broader peaks. The technique was successfully used to screen microgram quantities of a panel of therapeutic antibodies to identify candidates with low solubility in PBS.

Conclusions

The cross-interaction chromatography methods described can be used to screen large panels of recombinant antibodies in order to discover those with low solubility. Addition of this tool to the array of tools available for characterization of affinity and activity of antibody therapeutic candidates will improve selection of candidates with biophysical properties favorable to development of high concentration antibody formulations.

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Abbreviations

CIC:

cross-interaction chromatography

Fab:

fragment antigen binding

mAb:

monoclonal antibody

PBS:

phosphate buffered saline

pIgG:

polyclonal immunoglobulin

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Acknowledgements

We thank Dr. Charles Henry and Dr. Wilbur Wilson for helpful discussions and the members of Centocor Discovery Research for protein expression and purification.

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Authors and Affiliations

  1. Discovery Research, Centocor R&D, Radnor, Pennsylvania, 19087, USA

    Steven A. Jacobs, Sheng-Jiun Wu, Yiqing Feng, Deidra Bethea & Karyn T. O’Neil

  2. 145 King of Prussia Rd., Radnor, Pennsylvania, 19087, USA

    Steven A. Jacobs

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  1. Steven A. Jacobs
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  2. Sheng-Jiun Wu
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Correspondence to Steven A. Jacobs.

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Jacobs, S.A., Wu, SJ., Feng, Y. et al. Cross-Interaction Chromatography: A Rapid Method to Identify Highly Soluble Monoclonal Antibody Candidates. Pharm Res 27, 65–71 (2010). https://doi.org/10.1007/s11095-009-0007-z

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  • DOI: https://doi.org/10.1007/s11095-009-0007-z

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