title: Chip-based cartilage oligomeric matrix protein detection in serum and synovial fluid for osteoarthritis diagnosis.
abstract: We have developed a method to detect cartilage oligomeric matrix protein (COMP) as a specific biomarker of osteoarthritis (OA). In pathological conditions of the cartilage, COMP is released first into the synovial fluid (SF) and from there into the blood. Thus, measurement of COMP in the blood and SF facilitates OA diagnosis. To determine COMP, we developed a fluoro-microbead guiding chip (FMGC)-based immunoassay. The FMGC has four immunoreactive regions, each with five patterns, to allow multiple assays. A COMP-specific capture antibody was immobilized to the FMGC surface to create a self-assembled interfacial layer. SF or serum samples from patients with OA possessing the target COMP were applied to the COMP-sensing monolayer. To generate binding signal, COMP detection antibody-conjugated fluoro-microbeads were applied and the numbers of fluoro-microbeads bound specifically were counted to determine COMP concentrations. This FMGC-based immunoassay clearly distinguished immunospecific from nonspecific binding by comparing optical signals from inside and outside of the patterns. The optical signals showed linear correlations with serum and SF COMP concentrations. Optical detection and quantification of COMP using fluorescence microscopy correlated well with results from commercial enzyme-linked immunosorbent assay (ELISA). This FMGC-based immunoassay offers a new approach for detecting a clinically relevant biomarker for OA in human blood and SF.
authors: Department of Molecular Science and Technology, Ajou University, Suwon 443749, Republic of Korea.
authors: Seung Yeon
authors: SY
authors: Song
authors: Yong Duk
authors: YD
authors: Han
authors: Sung Yub
authors: SY
authors: Hong
authors: Kangil
authors: K
authors: Kim
authors: Sang Sik
authors: SS
authors: Yang
authors: Byoung-Hyun
authors: BH
authors: Min
authors: Hyun C
authors: HC
authors: Yoon
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology; D064236:Cartilage Oligomeric Matrix Protein; D002417:Cattle; D016326:Extracellular Matrix Proteins / Q000032:analysis* / Q000097:blood / Q000276:immunology; D006023:Glycoproteins / Q000032:analysis* / Q000097:blood / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D056656:Lab-On-A-Chip Devices; D008297:Male; D064235:Matrilin Proteins; D008863:Microspheres; D055620:Optical Phenomena; D010003:Osteoarthritis / Q000097:blood / Q000175:diagnosis*; D013582:Synovial Fluid / Q000737:chemistry*
chemical_list: D055910:Antibodies, Immobilized; D064236:Cartilage Oligomeric Matrix Protein; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D064235:Matrilin Proteins; C577553:TSP5 protein, human