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Aptamer selection and aptasensor construction for bone density biomarkers.

Raja Chinnappan1, Norhan Sameh Zaghloul2, Razan AlZabn1

  • 1Department of Chemistry, Alfaisal University, Al Zahrawi Street, Al Maather, Al Takhassusi Rd, Riyadh, 11533, Saudi Arabia.

Talanta
|December 31, 2020
PubMed
Summary

New aptasensors offer a sensitive and cost-effective method for detecting osteoporosis biomarkers osteocalcin (OC) and beta-crosslap (BC). These aptamer-based biosensors overcome limitations of traditional antibody assays, enabling better monitoring of bone disease and therapeutic responses.

Keywords:
AptamerAptasensorBeta-crosslapBiosensorFRETGraphene-oxideOsteocalcinOsteoporosis

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Area of Science:

  • Biomarker Discovery
  • Biosensor Technology
  • Biochemistry

Background:

  • Osteoporosis (OP) is characterized by bone metabolism dysregulation, leading to porous bones.
  • Osteocalcin (OC) and beta-crosslap (BC) are key biomarkers for bone formation and resorption, respectively.
  • Current antibody-based assays for OC and BC have limitations including thermostability, sensitivity, animal use, and cost.

Purpose of the Study:

  • To develop novel aptamer-based assays for sensitive and specific detection of OC and BC.
  • To overcome the limitations associated with traditional antibody-based detection methods for osteoporosis biomarkers.
  • To fabricate and validate graphene oxide-based aptasensors for OC and BC detection.

Main Methods:

  • Systematic evolution of ligands by exponential enrichment (SELEX) was used to select specific DNA aptamers for OC and BC.
  • Binding affinity (Kd) of selected aptamers was evaluated against respective biomarkers.
  • Graphene oxide-based aptasensors were fabricated using high-affinity aptamers and tested with spiked buffer and rat serum samples.

Main Results:

  • High-affinity aptamers for OC and BC were successfully selected with Kd values of 59 nM and 55 nM, respectively.
  • Fabricated aptasensors achieved low detection limits of 0.4 pg/ml for OC and 0.21 pg/ml for BC.
  • The aptasensors demonstrated effective detection and validation in serum samples from osteoporotic rats.

Conclusions:

  • Aptamer-based assays, specifically graphene oxide aptasensors, provide a promising alternative for detecting osteoporosis biomarkers OC and BC.
  • These novel aptasensors offer high sensitivity, specificity, and overcome drawbacks of conventional antibody-based methods.
  • The developed aptasensors hold potential for improved diagnostics and monitoring of osteoporosis and related therapies.