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Tagging and Fusion Proteins

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

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Updated: Jun 10, 2025

Synthetic Methodology for Asymmetric Ferrocene Derived Bio-conjugate Systems via Solid Phase Resin-based Methodology

Synthetic Methodology for Asymmetric Ferrocene Derived Bio-conjugate Systems via Solid Phase Resin-based Methodology

Published on: March 12, 2015

HaloTag-Modified, Ferrocene Labeled Self-Assembled Monolayers for Protein Sensing.

Andrew R Brotherton¹, Guru P Neupane², Xudong Ji²

¹Department of Chemistry, 2145 Sheridan Road, Evanston, Illinois 60208-3113, United States.

Langmuir : the ACS Journal of Surfaces and Colloids

|October 16, 2024

Summary

This summary is machine-generated.

Ferrocene-based molecules in self-assembled monolayers (SAMs) show antifouling properties and specific detection of HaloTag protein. The position of ferrocene within the SAM influences nonspecific adsorption and electrochemical response to the protein.

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Area of Science:

Electrochemistry
Surface Science
Biomolecular Engineering

Background:

Self-assembled monolayers (SAMs) are crucial for modifying electrode surfaces.
Ferrocene (Fc) is a versatile redox reporter for electrochemical sensing.
Protein detection on surfaces requires strategies to minimize fouling and maximize specificity.

Purpose of the Study:

To investigate the antifouling and specific detection capabilities of Fc-based SAMs.
To explore the influence of Fc positioning within the SAM on protein interactions.
To demonstrate a proof-of-concept for electrochemical detection of HaloTag protein.

Main Methods:

Fabrication of mixed SAMs using Fc-based molecules and a PEG2 diluent.
Electrochemical characterization (cyclic voltammetry, capacitance measurements) before and after protein exposure.
Orthogonal validation using SAMs for MALDI mass spectrometry (SAMDI-MS).

Main Results:

Fc-SAMs exhibited antifouling properties, reducing nonspecific adsorption.
Specific detection of HaloTag protein was achieved, with Fc position influencing signal.
Increased disorder and ion mobility were observed upon protein binding.
SAMDI-MS confirmed HaloTag protein binding to the functionalized electrodes.

Conclusions:

Fc-based SAMs offer a promising platform for antifouling and specific protein detection.
Controlling the architecture of Fc-SAMs is key to optimizing sensor performance.
The study provides insights into designing electrochemical biosensors with enhanced selectivity and reduced fouling.