Electrochemical Biosensor: Point-of-Care for Early Detection of Bone Loss by Nasrin Afsarimanesh & Subhas Chandra Mukhopadhyay & Marlena Kruger

Author:Nasrin Afsarimanesh & Subhas Chandra Mukhopadhyay & Marlena Kruger
Language: eng
Format: epub
ISBN: 9783030037062
Publisher: Springer International Publishing

4.2 ELISA-Based Experiments

The initial experiments were performed using an ELISA kit in order to get idea about antigen-antibody-based methods. In addition, it was used to validate the results achieved from the developed sensor. As it was discussed in Chap. 2, most of the available methods for the assessment of biochemical markers of bone turnover are ELISA-based. ELISA is an accurate and reliable analytical tool that is generally used in biomedical research for the detection and quantification of a specific molecule in a liquid sample. ELISA can detect the analyte using enzyme-linked antigens and antibodies. Very small quantities of antigens such as hormones, proteins, peptides, or antibodies in a liquid sample can be detected and quantified using ELISA [6, 7].

The antigen in the liquid phase is immobilised into the wells of a 96-well micro-titre plate that links to a primary antibody. A secondary, enzyme-linked antibody then senses the antigen by binding the antigen to the antibody. A chromogenic substrate is used to vary colour in the presence of the antigen. Lastly, the measurement is completed using spectrophotometry technique [8]. Although this technique is a standard immunoassay method, there are some weaknesses in using ELISA; it is a laboratory-based assay that is time-consuming, costly and involves several steps and technical skill. It includes numerous steps and procedures for incubation, antibodies binding and measurements that require not only the services of highly skilled professionals and an expensive laboratory setup but also involves high costs for testing individual samples, and therefore cannot be used for frequent monitoring of CTx-I concentration to monitor variations in bone resorption in an individual.

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