Chaperone proteins as single component reagents to assess antibody nonspecificity.
MAbs
; 9(7): 1036-1040, 2017 10.
Article
in En
| MEDLINE
| ID: mdl-28745541
ABSTRACT
Early stage assays that evaluate monoclonal antibody drug-like properties serve as valuable tools for selection of lead candidates. One liability for clinical development, off-target reactivity, is often assessed by binding to a mixture or panel of noncognate proteins. While robust, these mixes are often ill-defined, and can suffer from issues such as lot-to-lot variability. In this study, we discovered in immunoprecipitation experiments that certain chaperones are present in one of these mixtures;we then explored the use of recombinant chaperone proteins as well-characterized agents to predict antibody nonspecificity. Antibody binding to the heat shock proteins HSP70, HSP90, or trigger factor all served as predictors of cross-interaction propensity, with HSP90 providing the greatest ability to predict antibody clearance rates in mouse. Individual chaperone binding correlates surprisingly closely with binding to complex cell extracts, with the exception of a few "false negatives" (assuming a complex cell extract as the "true" value). As defined reagents, these chaperone reagents present advantages for high throughput assays of nonspecificity.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Heat-Shock Proteins
/
Antibodies, Monoclonal
/
Antibody Specificity
Type of study:
Prognostic_studies
Limits:
Animals
/
Humans
Language:
En
Journal:
MAbs
Journal subject:
ALERGIA E IMUNOLOGIA
Year:
2017
Document type:
Article
Affiliation country:
Estados Unidos
Publication country:
EEUU
/
ESTADOS UNIDOS
/
ESTADOS UNIDOS DA AMERICA
/
EUA
/
UNITED STATES
/
UNITED STATES OF AMERICA
/
US
/
USA