Evaluation of Factors Influencing Antibody Reduction for Development of Antibody Drug Conjugates
Iran Biomed J
; 21(4): 270-4, 2017 07.
Article
in En
| MEDLINE
| ID: mdl-28410549
ABSTRACT
Background:
Reduction/alkylation is one of the leading strategies for the development of antibody drug conjugates (ADCs). Precise control of the reduction process would not only yield a defined number of free thiols per antibody but also result in development of more homogenous conjugates.Methods:
In the present study, we investigated the effect of various dithiothreitol (DTT) concentrations, temperature conditions, and DTT exposure times on antibody reduction. After antibody reduction, the Ellman's test and SDS-PAGE analysis were used to evaluate free thiols produced and confirm the reduction process, respectively.Results:
DTT concentration seems to be a potential factor in the reduction process. Concentrations of 0.1, 1, 5, 10, 20, 50, and 100 mM DTT at 37°C for 30 minutes resulted in approximately 0.4, 1.2, 5.4, 7, 8, 8, and 8 thiols per antibody, respectively.Conclusion:
Optimized site-specific conjugation can provide better process control and reproducibility for the development of disulfide-based ADCs.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
Iran Biomed J
Journal subject:
BIOLOGIA
/
MEDICINA
Year:
2017
Document type:
Article
Affiliation country:
Irán