Organo-Fonksiyonel Silan ile Modifiye Edilmiş ITO Elektrot Temelli CDH 22 Biyobelirteç Tespiti için Hassas ve Etiketsiz Bir Elektrokimyasal İmpedans İmmünosensörü

Bu çalışmada, anti-CDH 22 antikoru ve CDH 22 antijeni arasındaki spesifik immüno-reaksiyona dayanan, kadherin benzeri protein (CDH 22) biyobelirtecinin tespiti için yeni bir elektrokimyasal etiketsiz immünosensör üretilmiştir. Geliştirilen immünosensör, anti-CDH 22 antikorlarının 3-siyanopropiltrimetoksisilan (CPTMS) ile modifiye edilmiş ITO substratı üzerinde immobilizasyonu ile oluşturulmuştur. Anti-CDH 22 antikorlarının CPTMS silanizasyon ajanına etkili bağlanması, morfolojik karakterizasyon (Taramalı Elektron Mikroskobu (SEM), Atomik Kuvvet Mikroskobu (AFM)) ve elektrokimyasal karakterizasyon (Elektrokimyasal İmpedans Spektroskopisi (EIS), Siklik Voltammetri (CV) kullanılarak incelendi. Optimum deney koşulları altında, ITO / CPTMS ile modifiye edilmiş elektrot, anti-CDH 22 antikor bağlanması için iyi bir bağlayıcıydır. Ek olarak, CPTMS modifiye elektrot CDH 22 antijen tespiti için etkili bir yüzey sunmuştur. İmmünosensör, geniş bir lineer tespit aralığı (0.03-3 pg/mL) ile düşük tespit limiti (9 fg/mL) ile sahipti. Ayrıca, iyi tekrarlanabilirlik, mükemmel tekrarlanabilirlik ve uzun depolama kararlılığına sahipti. Ek olarak, önerilen immünosensörün pratik uygulanabilirliği, insan serum numuneleri kullanılarak araştırıldı. İnsan serum numuneleri geri kazanım sonuçları (% 99.16 -% 101.94) önerilen biyosensörün doğruluğunu göstermektedir. Sonuç olarak, CPTMS, biyosensör yapımı için umut verici bir platform olabilir ve bu önerilen immünosensör gerçek insan serum analizi için uygulanabilir.

A Sensitive and Label-Free Electrochemical Impedance Immunosensor for CDH 22 Biomarker Detection Based on Organo-Functional Silane Modified ITO Electrode

In this work, a novel electrochemical label free immunosensor was fabricated for Cadherin-like protein 22 (CDH 22) biomarker detection based on specific immunoreaction between anti-CDH 22 antibody and CDH 22 antigen. The developedimmunosensor was constructed through the immobilization of anti-CDH 22 antibodies on 3-cyanopropyltrimethoxysilane (CPTMS) modified ITO substrate. The effective binding of the anti-CDH 22 antibodies on the CPTMS silanization agentwas investigated by using morphological characterization (Scanning Electron Microscopy (SEM), Atomic Force Microscopy(AFM)) and electrochemical characterization (Electrochemical Impedance Spectroscopy (EIS), Cyclic Voltammetry (CV)).Under optimum experimental conditions, the ITO/CPTMS modified electrode was a good linker for anti-CDH 22 antibodyanchoring. In addition, CPTMS modified electrode offered an efficient surface to CDH 22 antigen detection. The immunosensor had a wide linear detection range (0.03-3 pg/mL) with low detection limit (9 fg/mL). Also, it had good reproducibility,excellent repeatability and long storage stability. In addition, the practical applicability of the proposed immunosensor wasinvestigated by utilizing human serum samples. The human serum samples recovery results (99.16% - 101.94%) illustratedthe accuracy of the suggested biosensor. Consequently, CPTMS can be a promising platform for biosensor construction andthis suggested immunosensor can be applicable for real human serum analysis.

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