HaloTag-based conjugation of proteins to barcoding-oligonucleotides.

ABSTRACT: Highly sensitive protein quantification enables the detection of a small number of protein molecules that serve as markers/triggers for various biological phenomena, such as cancer. Here, we describe the development of a highly sensitive protein quantification system called HaloTag protein barcoding. The method involves covalent linking of a target protein to a unique molecule counting oligonucleotide at a 1:1 conjugation ratio based on an azido-cycloalkyne click reaction. The sensitivity of the HaloTag-based barcoding was remarkably higher than that of a conventional luciferase assay. The HaloTag system was successfully validated by analyzing a set of protein-protein interactions, with the identification rate of 44% protein interactions between positive reference pairs reported in the lit

SUBMITTER: Yazaki J 

PROVIDER: S-EPMC6954424 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature