Cross-interaction chromatography: a rapid method to identify highly soluble monoclonal antibody candidates.
OBJECTIVE
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.
Jacobs
Steven A
SA
Discovery Research, Centocor R&D, Radnor, Pennsylvania 19087, USA. sjacobs9@its.jnj.com
Wu
Sheng-Jiun
SJ
Feng
Yiqing
Y
Bethea
Deidra
D
O'Neil
Karyn T
KT
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility; D014462:Ultrafiltration / Q000379:methods
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins