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

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...

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Bibliographic Details
Published in:Pharmaceutical research 2010, Vol.27 (1), p.65-71
Main Authors: Jacobs, Steven A., Wu, Sheng-Jiun, Feng, Yiqing, Bethea, Deidra, O’Neil, Karyn T.
Format: Article
Language:English
Subjects:
Antibodies, Monoclonal - chemistry
Biochemistry
Biological and medical sciences
Biomedical and Life Sciences
Biomedical Engineering and Bioengineering
Biomedicine
Chromatography
Chromatography, High Pressure Liquid - methods
Drug Evaluation, Preclinical - methods
General pharmacology
Humans
Medical Law
Medical sciences
Monoclonal antibodies
Pharmaceutical sciences
Pharmaceutical technology. Pharmaceutical industry
Pharmacology. Drug treatments
Pharmacology/Toxicology
Pharmacy
Proteins
Recombinant Proteins - chemistry
Research Paper
Solubility
Ultrafiltration - methods
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Summary: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.
ISSN:0724-8741
1573-904X
DOI:10.1007/s11095-009-0007-z