Protein G: β-galactosidase fusion protein for multi-modal bioanalytical applications

Biotechnol Prog. 2022 Aug 17;e3297. doi: 10.1002/btpr.3297. Online ahead of print.

Abstract

β-galactosidase (β-gal) is one of the most prevalent markers of gene expression. Its activity can be monitored via optical and fluorescence microscopy, electrochemistry, and many other ways after slight modification using protein engineering. Here, we have constructed a chimeric version that incorporates a streptococcal protein G domain at the N-terminus of β-gal that binds immunoglobins, namely IgG. This protein G: β-galactosidase fusion enables β-gal-based spectrophotometric and electrochemical measurements of IgG. Moreover, our results show linearity over an industrially relevant range. We demonstrate applicability with rapid spectroelectrochemical detection of IgG in several formats including using an electrochemical sensing interface that is rapidly assembled directly onto electrodes for incorporation into biohybrid devices. The fusion protein enables sensitive, linear, and rapid responses, and in our case, makes IgG measurements quite robust and simple, expanding the molecular diagnostics toolkit for biological measurement.

Keywords: beta-galactosidase; electrochemical assays; protein G; protein engineering.