Detecting features of antibody structure through their mediator-accessible redox activities

Area of Science:

• Biophysical Chemistry
• Analytical Chemistry
• Biotechnology

Background:

• Protein structure, including sequence, folding, and post-translational modifications, dictates function.
• Traditional molecular measurements are essential for characterizing protein structural features.
• Monitoring protein structural integrity is critical during biomanufacturing.

Purpose of the Study:

• To develop an alternative method for assessing protein structural features using electronic patterns.
• To validate an experimental approach for detecting structural perturbations in biomanufacturing.
• To demonstrate the utility of mediated electrochemical probing (MEP) for real-time bioprocess monitoring.

Main Methods:

• Utilized mediated electrochemical probing (MEP) to generate electronic patterns representing molecular features.
• Studied a NIST monoclonal antibody standard to detect structural variants.
• Focused on identifying interchain disulfide bond reduction and methionine oxidation.

Main Results:

• MEP successfully discerned electronic patterns associated with the antibody's mediator-accessible redox activity.
• The method rapidly (within minutes) and quantitatively detected structural alterations.
• Robust electronic signals were produced, indicating potential for biomanufacturing monitoring.

Conclusions:

• Mediated electrochemical probing offers a novel approach to transduce protein structural information into electronic signals.
• MEP can rapidly and quantitatively detect critical structural perturbations relevant to biomanufacturing.
• This electronic detection method enables the application of microelectronics and real-time data analytics to protein analysis.