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A quantitative lateral flow immunoassay for the determination of cartilage oligomeric matrix protein as a marker of osteoarthritis

Hong, Sung Yub; Park, Yoo Min; Jang, Yo Han; Min, Byoung-Hyun; Yoon, Hyun C.
Department of Orthopedic Surgery, Ajou University School of Medicine
This study reports a simple and quantitative analytical method for cartilage oligomeric matrix protein (COMP) using a lateral flow immunoassay. An immunochromatographic assay using gold nanoparticle (AuNPs)-labeled monoclonal antibody (12C4) probe for the quantitative detection of COMP in human synovial fluid was developed. The detection antibody was conjugated with AuNPs (30 nm) as a detector, and the capture antibody (16F12) was immobilized within a multi-spot pattern on the test zone of the membrane strip. The COMP in human synovial fluid binds with AuNP-conjugated detection antibody and then COMP-Ab complex binds to the capture antibody. The immunoassay test can be analyzed by visualizing the test strip with a commercial scanner and an imaging program. The color density of the multi-spot pattern at the test zone was increased in proportion to the concentration of COMP. The obtained results from the developed immunoassay exhibited a dynamic COMP detection range between 0.6 μg/mL and 10 μg/mL. The detection limit of the proposed lateral flow immunoassay system was about 0.2 μg/mL. Results from the developed analysis system were compared with a conventional ELISA. Signal from the developed analysis system and ELISA showed a good correlation. The assay time of the quantitative test of COMP was less than 5 min. We expect that the proposed immunoassay method is suitable for the practical osteoarthritis diagnosis based on the quantification of COMP.
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