Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review

Hafiz Muhammad Husnain Azam

Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review

Abstract

Nucleic acid biosensors rely on sequence-specific recognition, yet analytical performance is strongly influenced by the interface on which capture probes are presented. This mini-review examines how immobilization chemistry and probe-layer architecture govern hybridization at solid-liquid interfaces. Common strategies, including thiol-gold attachment, carbodiimide-mediated covalent coupling, biotin-streptavidin affinity immobilization, click chemistry, polyadenine anchoring, and DNA nanostructures, are compared in terms of stability, orientation, spacing, and accessibility. Particular emphasis is placed on probe density, probe-surface distance, electrostatic effects, secondary structure, surface heterogeneity, and their consequences for kinetics, selectivity, and signal generation. Platform-specific implications are discussed for electrochemical, optical, nanoparticle-based, and bead-based biosensors. Across these systems, high probe loading does not necessarily produce efficient target capture because crowding and unfavorable surface interactions can reduce the fraction of hybridization-competent probes. Practical design principles are proposed for separating immobilization yield, functional probe availability, and transduced signal. Future progress will depend on quantitatively defined biointerfaces that integrate controlled probe presentation, antifouling behavior, microfluidic transport, multiplexing, and validation in realistic biological samples.

Keywords

Nucleic acid biosensors, Surface biofunctionalization, Probe immobilization, Surface hybridization, Probe density, Biointerfaces

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Details

Primary Language

English

Subjects

Sensor Technology

Journal Section

Review

Authors

Hafiz Muhammad Husnain Azam ^*
0009-0001-3702-2313
Germany

Publication Date

October 22, 2026

Submission Date

August 10, 2026

Acceptance Date

October 6, 2026

Published in Issue

Year 2026 Number: 2026-1

IZ

https://izlik.org/JA94CH42EP

APA

Azam, H. M. H. (2026). Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review. Turkish Journal of Sensors and Biosensors, 2026-1, 1-21. https://izlik.org/JA94CH42EP

AMA

1.Azam HMH. Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review. Turkish Journal of Sensors and Biosensors. 2026;(2026-1):1-21. https://izlik.org/JA94CH42EP

Chicago

Azam, Hafiz Muhammad Husnain. 2026. “Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review”. Turkish Journal of Sensors and Biosensors, no. 2026-1: 1-21. https://izlik.org/JA94CH42EP.

EndNote

Azam HMH (October 1, 2026) Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review. Turkish Journal of Sensors and Biosensors 2026-1 1–21.

IEEE

[1]H. M. H. Azam, “Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review”, Turkish Journal of Sensors and Biosensors, no. 2026-1, pp. 1–21, Oct. 2026, [Online]. Available: https://izlik.org/JA94CH42EP

ISNAD

Azam, Hafiz Muhammad Husnain. “Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review”. Turkish Journal of Sensors and Biosensors. 2026-1 (October 1, 2026): 1-21. https://izlik.org/JA94CH42EP.

JAMA

1.Azam HMH. Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review. Turkish Journal of Sensors and Biosensors. 2026;:1–21.

MLA

Azam, Hafiz Muhammad Husnain. “Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review”. Turkish Journal of Sensors and Biosensors, no. 2026-1, Oct. 2026, pp. 1-21, https://izlik.org/JA94CH42EP.

Vancouver

1.Hafiz Muhammad Husnain Azam. Surface Biofunctionalization of Capture Oligonucleotides in Solid-Phase Nucleic Acid Hybridization Biosensors: A Critical Review. Turkish Journal of Sensors and Biosensors [Internet]. 2026 Oct. 1;(2026-1):1-21. Available from: https://izlik.org/JA94CH42EP

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