New insights on critical reagent optimization for antidrug antibody assays.
Bioanalysis
; 11(9): 815-823, 2019 May.
Article
em En
| MEDLINE
| ID: mdl-30974951
ABSTRACT
Aim:
Conjugated critical reagents are a pillar of ligand binding analysis. Although good practices for characterization have already been discussed, little is known about how assays are affected by specific conjugation parameters.Results:
Here we developed, characterized and screened a toolset of bioconjugates that provided new insights about the optimization of conjugated critical reagent attributes. Biotinylated and sulfo-tagged trastuzumab were utilized as capture and detection antibodies, respectively, in an antidrug antibody (ADA) assay. The optimal conjugation window was identified by functional assessment. Excess of unlabeled biotin, but not sulfo-tag, affected the assay performance. An increase in the assay baseline was observed when sulfo-tagged trastuzumab underwent increasing freeze-thaw cycles.Conclusion:
Upfront systematic screening and characterization of conjugated critical reagent attributes benefit assay robustness.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Imunoensaio
/
Trastuzumab
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article