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Published on: June 14, 2018
Monoclonal antibody selection for interleukin-4 quantification using suspension arrays and forward-phase protein
1Biochemical Science Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899-8312, USA. lili.wang@nist.gov
Journal of Proteome Research
|November 14, 2007
Summary
This study identified optimal antibody pairs for sensitive detection of interleukin-4 (IL-4) using high-throughput methods. Multiplexed bead arrays and protein microarrays are effective for selecting antibodies in systems biology and diagnostics.
Area of Science:
- Immunotechnology
- Biotechnology
- Assay Development
Background:
- Accurate detection of cytokines like interleukin-4 (IL-4) is crucial for biological research and clinical diagnostics.
- Selecting optimal monoclonal antibodies (mAbs) for sensitive immunoassays, such as sandwich assays, is essential for reliable measurements.
Purpose of the Study:
- To evaluate and compare the effectiveness of different monoclonal antibodies (mAbs) against mouse IL-4.
- To identify the most sensitive antibody pairs for detecting recombinant IL-4 using both conventional and high-throughput techniques.
Main Methods:
- Utilized a model system with recombinant mouse IL-4 and purified rat anti-mouse IL-4 mAbs.
- Employed surface plasmon resonance (SPR), suspension arrays (multiplexed bead arrays), and forward-phase protein microarrays for antibody selection.
- Performed sandwich-type immunoassay validation.
Main Results:
- SPR, suspension arrays, and protein microarrays consistently identified the same capture (BVD4-1D11) and detection (BVD6-24G2) antibody pair.
- This optimal antibody pair enabled detection of IL-4 down to 2 pg/mL in buffer and 13.5 pg/mL in mouse serum using multiplexed bead arrays.
- High-throughput methods proved suitable for selecting multiple antibodies against a single analyte.
Conclusions:
- Multiplexed bead arrays and protein microarrays are efficient alternatives to SPR for antibody selection, especially for large-scale screening.
- These high-throughput methods support the development of sensitive immunoassays for systems biology and reproducible clinical diagnostics.
- The identified antibody pair offers enhanced sensitivity for IL-4 detection, improving patient care through better diagnostics.
