NanoLuc Luciferase - A Multifunctional Tool for High Throughput Antibody Screening

Area of Science:

• Biotechnology
• Molecular Biology
• Immunology

Background:

• NanoLuc luciferase (Nluc) is a small, stable, ATP-independent bioluminescent protein.
• Existing antibody screening methods can be complex and require purification of antibody fragments.

Purpose of the Study:

• To develop a robust, ultra-high sensitivity screening system for therapeutic antibodies and fragments.
• To leverage Nluc for versatile antibody characterization and quantification without purification.

Main Methods:

• Utilized NanoLuc luciferase (Nluc) protein fusions.
• Developed cellular and solid-phase ELISA assays.
• Implemented homogeneous BRET-based screening assays.
• Performed relative affinity determinations via competition ELISA.
• Applied direct Western blotting techniques.

Main Results:

• Achieved ultra-high sensitivity for characterizing and quantifying antibody hits.
• Demonstrated versatility across multiple assay formats (ELISA, BRET, Western blot).
• Enabled screening without the need for antibody purification.

Conclusions:

• The Nluc-based system offers a highly robust and versatile solution for antibody drug screening.
• This purification-free approach significantly enhances efficiency in high-throughput screening.
• The Nluc fusion technology serves as a valuable tool for current and future antibody discovery pipelines.