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Intermolecular forces (IMF) are electrostatic attractions arising from charge-charge interactions between molecules. The strength of the intermolecular force is influenced by the distance of separation between molecules. The forces significantly affect the interactions in solids and liquids, where the molecules are close together. In gases, IMFs become important only under high-pressure conditions (due to the proximity of gas molecules). Intermolecular forces dictate the physical properties of...
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Related Experiment Video

Updated: Feb 1, 2026

Demonstration of Heterologous Complexes formed by Golgi-Resident Type III Membrane Proteins using Split Luciferase Complementation Assay
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Ratiometric Bioluminescent Sensor Proteins Based on Intramolecular Split Luciferase Complementation.

Yan Ni1, Remco Arts1, Maarten Merkx1

  • 1Laboratory of Chemical Biology and Institute for Complex Molecular Systems, Department of Biomedical Engineering , Eindhoven University of Technology , Den Dolech 2 , 5612 AZ Eindhoven , The Netherlands.

ACS Sensors
|December 12, 2018
PubMed
Summary

Researchers developed novel bioluminescent sensor proteins using split NanoLuc luciferase for sensitive antibody detection. This new method offers improved dynamic response for applications like HIV testing and therapeutic antibody quantification.

Keywords:
BRETantibodiesbiosensorsprotein engineeringsplit luciferase

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Bioluminescent sensor proteins are valuable tools for in vivo imaging and point-of-care diagnostics.
  • Existing sensors often rely on direct modulation of bioluminescence resonance energy transfer (BRET) efficiency.
  • There is a need for novel sensor designs with enhanced sensitivity and dynamic range.

Purpose of the Study:

  • To introduce a new class of ratiometric bioluminescent sensor proteins based on competitive intramolecular complementation of split NanoLuc luciferase.
  • To demonstrate the feasibility of this sensor principle through proof-of-concept studies.
  • To showcase the modularity and applicability of the sensor architecture for rational engineering.

Main Methods:

  • Development of a blue-red light emitting sensor protein utilizing competitive intramolecular complementation of split NanoLuc luciferase.
  • Application of the sensor for the detection of anti-HIV1-p17 antibodies.
  • Testing the sensor's performance for the quantification of the therapeutic antibody cetuximab in undiluted blood plasma.

Main Results:

  • A novel sensor protein achieved a 500% change in emission ratio and a Kd of 10 pM for anti-HIV1-p17 antibodies.
  • The sensor design improved the dynamic response of a cetuximab sensor by 4-fold.
  • Direct quantification of cetuximab in undiluted blood plasma was enabled.

Conclusions:

  • A new principle for ratiometric bioluminescent sensors based on split luciferase complementation has been established.
  • The developed sensors demonstrate high sensitivity and improved dynamic response for antibody detection.
  • The modular sensor architecture facilitates systematic tuning and rational engineering of bioluminescent sensors for diverse applications.