Quantitative lateral-flow immunoassay for the assessment of the cartilage oligomeric matrix protein as a marker of osteoarthritis

Abstract

This study reports a simple and quantitative analytical method for detecting cartilage oligomeric matrix protein (COMP) using a lateral-flow immunoassay (LFI) format. An immuno-chromatographic assay was developed using a gold nanoparticle (AuNP)-conjugated monoclonal antibody (Ab) probe for the quantitative detection of COMP in human synovial fluid (SF). The detection antibody was conjugated with the AuNP (~30 nm) to enable detection, and the capture antibody was immobilized within a multi-spot pattern in the test zone of a membrane strip. COMP in the human SF binds to the AuNP-conjugated detection antibody, and the COMP-Ab complex then binds to the capture antibody. The immunoassay test can be analyzed by visualizing the test strip with a commercial scanner and an imaging software program. The color density of the multi-spot pattern at the test zone increased in proportion to the concentration of COMP. The developed immunoassay exhibited a dynamic detection range from 0.6 to 20 μg/mL of COMP. The detection limit of the proposed LFI system was approximately 0.2 μg/mL. The test results showed good correlation with those obtained using a conventional enzyme-linked immunosorbent assay (ELISA) to detect COMP. We expect that the proposed immunoassay method, based on the quantification of COMP, is suitable for osteoarthritis diagnosis using SF samples.

Nanoparticle-based lateral flow biosensors

“…In the work of Hong et al (2012) the antibodies were dispensed as patterns of parallel and zigzag dots on the detection pad, being the zigzag pattern the one which gave the highest intensities, presumably being flow slower on this design. Recently, it has been demonstrated that wax is a useful tool due its hydrophobicity to control flow on paper devices and create channels on it (Renault et al, 2014).…”

Emerging Technologies and Platforms for the Immunodetection of Multiple Biochemical Markers in Osteoarthritis Research and Therapy

“…For instance, a quantitative lateral-flow immunoassay technique with antibodies conjugated to gold nanoparticles was used for the detection of COMP in the synovial fluid. The proposed method showed similar results to corresponding ELISA, with an average difference of <7% without the need of expensive equipment or complex procedures (127). However, most of the reported studies have investigated the use of advanced nanomaterial-enhanced immunoassays toward inflammatory biochemical markers.…”

Instrumentation-Free Semiquantitative Immunoanalysis Using a Specially Patterned Lateral Flow Assay Device

“…The 20 nm AuNPs were synthesized according to the procedure described in previous reports [ 32 , 33 ]. The size of the AuNPs was determined with an ultraviolet–visible (UV–vis) spectrophotometer by scanning from 300 to 800 nm ( Figure S1A, Supporting information ) [ 34 ].…”

“…The distributed spots allow for the quantitative analysis of an LFI with the naked eye. However, the accumulated immunocomplexes on the spots could affect the flow on the strip because of their large components, such as antigens, antibodies, and AuNPs [33]. The color development of the spot is not only determined by the concentration of antigens but also by the contact chance between the flow and spot.…”