Membrane-Based Microwave-Mediated Electrochemical Immunoassay for the In Vitro, Highly Sensitive Detection of Osteoporosis-Related Biomarkers

Abstract

Highly sensitive and multiplexed in vitro detection of osteoporosis-related biochemical markers were carried out based on the membrane-based microwave-mediated electrochemical immunoassay (MMeEIA), where we can dramatically reduce the sample preparation time by shortening the incubation time of conjugation to obtain sensitive detection based on three dimensional conjugation of antibodies with target antigens in nylon membrane disk. C-terminal cross-linked telopeptide of type I collagen (CTx), Osteocalcin (OC), parathyroid hormone (PTH), and N-terminal propeptide of type I collagen (P1NP), which can be utilized to monitor the progress of osteoporosis, were quantified using their corresponding antibody immobilized in membranes. Coefficient of variations in this intra- and inter-assays were within 8.0% for all markers. When compared with data obtained from clinically used standard equipment (Roche modular E170), their coefficients of determination, R2 values, are mostly more than 0.9. They show that the results obtained from MMeEIA are in good agreement with that from the conventional clinical instruments.

Recent research trends in voltammetric sensing platforms for hormones and their applications to human serum analyses

“…Another peptide hormone, PTH, which generally controls the metabolism of calcium and phosphate in the blood, is related to osteoporosis and parathyroid cancer. 55,58 A voltammetric sensor was recently reported for the analysis of PTH in human serum via the immobilization of horseradish peroxidase (HRP) labeled anti-PTH on an AuNPs and SWCNTs modified SPCE. 59 In that work, the role of AuNPs and SWCNTs was an immobilization scaffold.…”

Multiplexed Immunosensors and Immunoarrays

“…Clinically relevant LODs of 1 pg mL –1 for insulin and millimolar levels for glucose were achieved, and direct measurements in blood and saliva were demonstrated. Kim et al employed membrane-based microwave-mediated electrochemical immunoassay based on fast conjugation of antibodies with target antigens in a three-dimensional nylon membrane to detect osteoporosis biomarkers including type I collagen peptides, Osteocalcin (OC), and parathyroid hormone (PTH) . Microwave radiation was used here to greatly shorten incubation time for efficient antibody–antigen binding.…”

“…Kim et al employed membrane-based microwave-mediated electrochemical immunoassay based on fast conjugation of antibodies with target antigens in a three-dimensional nylon membrane to detect osteoporosis biomarkers including type I collagen peptides, Osteocalcin (OC), and parathyroid hormone (PTH). 29 Microwave radiation was used here to greatly shorten incubation time for efficient antibody−antigen binding. Detection employed an alkaline phosphatase label that produced p-aminophenol for differential pulse voltammetry measurement by a separate sensor electrode.…”