1 research outputs found
Analytical Cascades of Enzymes for Sensitive Detection of Structural Variations in Protein Samples
Protein
function critically depends on structure. However, current
analytical tools to monitor consistent higher-order structure with
high sensitivity, as for instance required in the development of biopharmaceuticals,
are limited. To complement existing assays, we present the analytical
cascade of enzymes (ACE), a method based on enzymatic modifications
of target proteins, which serve to exponentially amplify structural
differences between them. The method enables conformational and chemical
fingerprinting of closely related proteins, allowing for the sensitive
detection of heterogeneities in protein preparations with high precision.
Using this method, we detect protein variants differing in conformation
only, as well as structural changes induced by diverse covalent modifications.
Additionally, we employ this method to identify the nature of structural
variants. Moreover, the ACE method should help to address the limited
reproducibility in fundamental research, which partly relates to sample
heterogeneities