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A novel monoclonal antibody screening method using the Luminex-100 microsphere system
Jonathan Seideman1, David Peritt
1Department of Immunobiology, Centocor, 200 Great Valley Parkway, Malvern, PA 19355, USA. jseidema@cntus.jnj.com
Journal of Immunological Methods
|August 8, 2002
Summary
We developed a sensitive Luminex assay to screen for neutralizing antibodies against interleukin-6 (IL-6). This high-throughput screening method efficiently detects antibodies, aiding in the development of new therapeutics.
Area of Science:
- Biotechnology
- Immunology
- Assay Development
Background:
- Neutralizing antibodies are crucial for targeted therapies.
- Developing efficient screening methods for these antibodies is essential.
Purpose of the Study:
- To develop a robust and sensitive assay for screening neutralizing antibodies.
- To demonstrate the assay's effectiveness using interleukin-6 (IL-6) as a model.
Main Methods:
- Utilized Luminex-100 microspheres coupled with a soluble IL-6 receptor.
- Employed biotin-labeled IL-6 as a probe for binding competition assays.
- Tested the assay's sensitivity with three anti-IL-6 monoclonal antibodies.
Main Results:
- Achieved a detection limit of <500 pg/ml for biotin-IL-6.
- Demonstrated the ability to detect antibody concentrations as low as 10 ng/ml.
- Confirmed assay robustness without interference from hybridoma components.
Conclusions:
- The developed Luminex-based assay is a sensitive and robust system for screening neutralizing antibodies.
- This high-throughput screening (HTS) method is time-efficient and suitable for antibody discovery.
- The assay is applicable whenever a soluble receptor for a target molecule is available.