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A modified ELISA assay differentiates CCL20 locked dimers from wild-type monomers
Xuesong Wu1, William R Clarke2, Chad A Koplinski3
1Department of Dermatology, University of California, Davis, Sacramento, CA, USA.
Journal of Immunological Methods
|March 2, 2023
Summary
A new assay specifically detects engineered CCL20 locked dimer (CCL20LD), a potential psoriasis treatment. This method is crucial for evaluating CCL20LD
Area of Science:
- Biochemistry
- Immunology
- Pharmacology
Background:
- Psoriasis and psoriatic arthritis are inflammatory diseases.
- Engineered CCL20 locked dimer (CCL20LD) is a novel therapeutic candidate targeting CCR6-mediated chemotaxis.
- Accurate quantification of CCL20LD serum levels is essential for preclinical drug development.
Purpose of the Study:
- To develop a novel immunoassay for specific quantification of CCL20LD.
- To differentiate CCL20LD from wild-type CCL20 (CCL20WT) in biological samples.
- To validate the assay for preclinical assessment of CCL20LD pharmacokinetics.
Main Methods:
- Screening of monoclonal antibodies for specific CCL20LD recognition.
- Development of a sandwich ELISA using a selected antibody pair (capture and biotinylated detection).
- Validation with recombinant proteins and analysis of serum samples from treated mice.
Main Results:
- Identification of a monoclonal antibody clone suitable for both capture and detection.
- Establishment of a CCL20LD-selective ELISA assay with high specificity.
- Demonstration of assay utility in analyzing serum from CCL20LD-treated mice.
Conclusions:
- A novel, specific ELISA assay for CCL20LD quantification has been developed.
- This assay is suitable for preclinical studies of CCL20LD as a therapeutic for psoriatic diseases.
- The assay facilitates pharmacokinetic and drug evaluation for CCL20LD biopharmaceutical development.
