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

Motabar, Dana; Wang, Sally; Tsao, Chen-Yu; Payne, Gregory F; Bentley, William E

Motabar, Dana; Wang, Sally; Tsao, Chen-Yu; Payne, Gregory F; Bentley, William E.

Afiliación

Motabar D; Fischell Department of Bioengineering, University of Maryland, College Park, MD, USA.
Wang S; Institute for Bioscience and Biotechnology Research, University of Maryland, College Park, MD, USA.
Tsao CY; Robert E. Fischell Institute for Biomedical Devices, University of Maryland, College Park, MD, USA.
Payne GF; Fischell Department of Bioengineering, University of Maryland, College Park, MD, USA.
Bentley WE; Institute for Bioscience and Biotechnology Research, University of Maryland, College Park, MD, USA.

Biotechnol Prog ; 38(6): e3297, 2022 11.

Article en En | MEDLINE | ID: mdl-35976745

RESUMEN

ß-galactosidase (ß-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 ß-gal that binds immunoglobins, namely IgG. This protein G: ß-galactosidase fusion enables ß-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.

Asunto(s)

Inmunoglobulina G; Ingeniería de Proteínas; beta-Galactosidasa/química; Inmunoglobulina G/genética

Palabras clave

beta-galactosidase; electrochemical assays; protein G; protein engineering

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Inmunoglobulina G / Ingeniería de Proteínas Idioma: En Revista: Biotechnol Prog Asunto de la revista: BIOTECNOLOGIA Año: 2022 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

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