Nimet YILDIRIM TİRGİL, Yasin FEDAYİ, Zennure AÇIKGÖZ

BİYOMARKERLERİN İYİLEŞTİRİLMİŞ TESPİTİ İÇİN KARBON NANOTÜPLERLE MODİFİYE EDİLMİŞ ELISA SİSTEMİNİN GELİŞTİRİLMESİ

Bu çalışmada, bir peptit biyobelirtecini (İnsan VEGF) analiz etmek için nanomateryalle modifiye edilmiş bir ELISA sistemi geliştirdi ve değerlendirdi. Çok duvarlı karbon nanotüpler (MWCNT), yakalama/tespit antikorları ve HRP enzimi için nanomateryal bazlı modifiye malzemesi olarak kullanıldı. ELISA yönteminin bu üç farklı bağlanma aşamasındaki CNT modifikasyonları test edildi ve karşılaştırıldı. Bu çalışmaların hassasiyet, doğrusallık ve tekrarlanabilirliklerine bağlı olarak en verimli modifikasyon adımı belirlendi. CNT'lerin kullanıldığı ELISA metodunda, özellikle 0,5 pg/ml tespit limit değeri ile daha yüksek hassasiyetler ve dolayısıyla daha güvenilir sonuçlar elde edildiği ve geleneksel ELISA yönteminede performansı artırmak için başarılı bir şekilde kullanılabileceğini kanıtlandı. Ayrıca ticari serum örneklerinin analizinde farklı modifikasyon adımlarının performansları ve tüm bu bağlanma adımlarının stabilitesi değerlendirildi ve geleneksel ELISA yöntemi ile karşılaştırıldı.

Anahtar Kelimeler:

ELISA, Karbon nanotüp, Biyoanaliz, Biyoteknoloji

CARBON NANOTUBES MODIFIED ELISA SYSTEM DEVELOPMENT FOR IMPROVED DETECTION OF BIOMARKERS

This study developed and evaluated a nanomaterial-modified ELISA system to analyze a peptide biomarker (Human VEGF). Multiwalled carbon nanotubes (MWCNTs) were used as nanomaterial-based modifiers for the capture/detection antibodies and HRP enzyme. CNTs modifications of the ELISA method at these three different binding steps were tested and compared. The most efficient modification step was concluded depending on these works’ sensitivities, linearities, and repeatabilities. We proved that CNTs could be used successfully to enhance the performances compared to the traditional ELISA method by achieving higher sensitivities and, therefore, more confident results, especially with a 0.5 pg/ml of limit of detection value. In addition, the performances of different modification steps in the commercial serum sample and the stability of all these binding steps were evaluated, and compared with the traditional ELISA method.

Keywords:

ELISA, Carbon nanotube, Bioanalysis, Biotechnology,

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