Carbon nanofiber nanomaterial modified polystyrene enzyme linked immunosorbent assay substrate for detecting osteoporosis

Abstract

Osteoporosis (OS) develops without any pain or symptom, and it is usually identified when the weakened bone causes fracture. To overcome this issue, measuring bone density (BMD) helps to identify osteoporosis and its conditions. It was identified that when the level of anti-cyclic citrullinated peptide (anti-CCP) increases, values of BMD were decreased. This research was focused to quantify the level of anti-CPP on nanomaterial modified polystyrene ELISA substrate. ELISA wells were modified into amine-carbon nanofiber and then glutaraldehyde used as a linker to attach CCP. Anti-CCP antibody level was quantified on PS plate by secondary-antibody conjugated horseradish peroxidase. Carbon nanofiber improves the immobilization of CCP on PS well and attract higher number of anti-CCP antibody, which helps to reach the limit of detection at 10 pg/mL, which was four times lower limit of detection with conventional ELISA (40 pg/mL). Further, specific experiment was conducted with serum spiked anti-CCP antibody shows the absorbance increment in all the concentrations of anti-CCP antibody and control performances with relevant molecules did not increase the absorbance level confirms the selective and specific detection. This carbon nanofiber modified ELISA well quantifies anti-CCP level and helps to identify the condition of bone for diagnosing osteoporosis.