ArticleAnalytical and bioanalytical chemistry2025
A one-pot purification-free strategy for biotinylated antibody preparation enables sensitive ELISA detection of cardiac troponin I-C complex.
Article in Analytical and bioanalytical chemistry, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.
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Abstract
Biotinylated antibodies are widely used in ELISA. However, their preparation typically requires labor-intensive purification steps to remove the excess biotinylating reagent. Here, we present a novel one-pot, purification-free strategy for preparing biotinylated antibodies, enabling a robust sandwich ELISA platform for the detection of the cardiac troponin I-C complex, a key biomarker for acute myocardial infarction. By introducing glycine (Gly) as a quenching agent to inactivate the residual biotinylating reagent (NHS-biotin esters) during the labeling process, our method eliminates the need for post-labeling purification while effectively reducing nonspecific background. Systematic optimization of labeling conditions yielded an assay with a detection limit of 1.08 pM and a broad linear quantification range from 3.6 pM to 1 nM (R
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