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Controlled-potential coulometry, also known as potentiostatic coulometry, employs a three-electrode system in which the working electrode's potential is precisely regulated using a potentiostat. Platinum working electrodes are utilized for positive potentials, while mercury pool electrodes are favored for extremely negative potentials. The platinum counter electrode is separated from the analyte using a membrane or salt bridge to avoid interference in the analysis.
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Related Experiment Video

Updated: May 29, 2025

Selection of Transporter-Targeted Inhibitory Nanobodies by Solid-Supported-Membrane SSM-Based Electrophysiology

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Published on: May 3, 2021

Everything's under Control: Maximizing Biosensor Performance through Negative Control Probe Selection.

Joseph Bucukovski¹, Benjamin L Miller^1,2,3,4,5

¹Department of Biochemistry and Biophysics, University of Rochester, Rochester, New York 14627, United States.

Analytical Chemistry

|February 3, 2025

Summary

This summary is machine-generated.

Selecting the right negative control probe is crucial for accurate label-free biosensing. This study presents a framework to optimize negative controls, showing the best choice varies by analyte for reliable disease monitoring and drug discovery.

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Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

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Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

Published on: April 17, 2017

Area of Science:

Biomedical Engineering
Analytical Chemistry
Biotechnology

Background:

Label-free biosensing enables macromolecule detection in complex biological samples for disease monitoring and drug discovery.
Nonspecific binding in complex matrices like serum is a major challenge, obscuring specific binding signals.
Distinguishing specific from nonspecific binding in label-free biosensors necessitates a reference (negative control) probe for signal subtraction.

Purpose of the Study:

To develop an FDA-inspired framework for selecting optimal negative control probes in label-free biosensing.
To systematically analyze and determine the best negative control probe for specific analytes using photonic ring resonator sensors.
To evaluate the impact of negative control selection on assay performance metrics like linearity, accuracy, and selectivity.

Main Methods:

Utilized photonic ring resonator sensors with two distinct monoclonal antibody capture probes.
Implemented an FDA-inspired framework for systematic evaluation of potential negative control probes.
Assessed assay performance using bioanalytical parameters including linearity, accuracy, and selectivity for two analytes: IL-17A and CRP.

Main Results:

The optimal negative control probe differed for IL-17A and CRP, despite subtle differences in assay performance.
For IL-17A detection, BSA achieved the highest score (83%), followed closely by mouse IgG1 isotype control (75%).
For CRP detection, rat IgG1 isotype control scored highest (95%), with anti-FITC as the second best (89%).

Conclusions:

The selection of an optimal on-chip reference control probe is analyte-dependent and requires case-by-case optimization.
Isotype-matching to the capture antibody is not always the best strategy for negative control selection.
The proposed framework provides a systematic approach to identify the most effective negative control for label-free biosensing assays.