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ToxSci Advance Access originally published online on December 22, 2003
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Toxicological Sciences 77, 280-289 (2004)
Copyright © 2004 by the Society of Toxicology

IMMUNOTOXICOLOGY

An In Vitro Human Cell–Based Assay to Rank the Relative Immunogenicity of Proteins

Marcia Stickler*, Narapon Rochanayon*, O. Jennifer Razo*, Jeanette Mucha*, Wendy Gebel*, Nargol Faravashi*, Regina Chin*, Susan Holmes{dagger} and Fiona A. Harding*,1

* Genencor International, Palo Alto, California 94304, and {dagger} Department of Statistics, Stanford University, Stanford, California 94305

A method to rank proteins based on their relative immunogenicity has been devised. A statistical analysis of peptide-specific responses in large human donor pools provides a structure index value metric that ranked four industrial enzymes in the order determined by both mouse and guinea pig exposure models. The ranking method also compared favorably with human sensitization rates measured in occupationally exposed workers. Structure index values for other proteins known to cause immune responses in humans were also determined and found to be higher than the value determined for human ß2-microglobulin. Using values from known immunogenic and putative nonimmunogenic proteins, a cut-off value was established. The structure index value calculation provides a comparative method to predict subsequent immunogenicity on a human population basis without the need to use animal models. Information provided by this assay can be used in the early development of protein therapies and other protein-based applications to select or create reduced immunogenicity variants.

Key Words: immunogenicity; cellular activation; human; antigen/peptide/epitopes.


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