Year 2021, Volume 11 , Issue 2, Pages 1279 - 1292 2021-06-01

Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi
Development of a New Disposable Label-free AFP Immunosensor for Electrochemical Detection of Cancer Biomarker Alpha-Fetoprotein

Kadir ERDEM [1] , Melike BİLGİ KAMAÇ [2]


Bu araştırmada, bir kanser biyobelirteci olan alfa-fetoproteinin (AFP) elektrokimyasal tayini için tek kullanımlık yeni etiketsiz immünosensör geliştirilmiş ve insan kan serumu numunelerinde uygulaması gerçekleştirilmiştir. Çalışmada yüzey baskılı karbon elektrotlar (SPCE) ilk olarak indirgenmiş grafen oksit (RGO) ile modifiye edilmiş ardından elektropolimerizasyon tekniği uygulanarak elektrot yüzeyi polinötral kırmızı (PNR) ile kaplanmış, son olarak elektrodepozisyon metodu ile altın nanoparçacıklar (AuNP) yüzeyde oluşturulmuştur. SPCE/RGO/PNR/AuNP elektrodunun elektrokimyasal karakterizasyonu dönüşümlü voltametri (CV), diferansiyel puls voltametri (DPV) ve kare dalga voltametri (SWV) metotları kullanılarak yapılmıştır. SPCE/RGO/PNR/AuNP elektrotların yüzeyine Anti-AFP’nin immobilizasyonu 3-merkaptopropiyonik asit (3-MPA) ve 1-etil-3-(3-dimetilamino-propil) karbodiimid/N-hidroksisüksinimid (EDC/NHS) kullanılarak kovalent bağlama yöntemi ile gerçekleştirilmiştir. Hazırlanan AFP immünosensörünün optimum çalışma parametreleri DPV ve SWV metotları ile belirlenmiştir. DPV ve SWV metotları ile belirlenen doğrusal tayin aralıkları 1-500 ng mL-1, tespit limitleri ise sırasıyla 0.79 ng mL-1 ve 0.86 ng mL-1 olarak belirlenmiştir. AFP immünosensörünün tekrarlanabilirliği 50 ng mL-1 AFP konsantrasyonu için test edilmiş ve bağıl standart sapma (R.S.D.) değerleri DPV ve SWV metotları için sırasıyla %4.06 (n=10) ve %3.68 (n=10) olarak hesaplanmıştır. Geliştirilen AFP immünosensörünün uygulama kararlılığı 60 gün boyunca, depolama kararlılığı ise 12 hafta depolama süresince voltametrik metotlarla incelenmiştir. Hazırlanan AFP immünosensörleri insan serumu örneklerine bilinen miktarda ilave edilen farklı konsantrasyonlardaki AFP’nin analizi için kullanılmış ve %95’in üstünde geri kazanımlar elde edilmiştir.
In this study, a new disposable label-free immunosensor was developed for the electrochemical determination of alpha-fetoprotein (AFP) that is a cancer biomarker and its application was performed on human blood serum samples. In the work, firstly the screen printed carbon electrodes (SPCE) were modified with reduced graphene oxide (RGO), then the electrode surface was coated with polyneutral red (PNR) by applying the electropolymerization technique and finally, gold nanoparticles (AuNP) were created by electrodeposition method on the surface. Electrochemical characterizations of the SPCE/RGO/PNR/AuNP electrode were performed using cyclic voltammetry (CV), differential pulse voltammetry (DPV), and square wave voltammetry (SWV) methods. Immobilization of Anti-AFP to the surface of SPCE/RGO/PNR/AuNP electrodes was carried out by a covalent binding method using 3-mercaptopropionic acid (3-MPA) and 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide/N-hydroxysuccinimide (EDC/NHS). The optimum working parameters of the prepared AFP immunosensor were determined by DPV and SWV methods. It was calculated that the linear range was 1-500 ng mL-1, the limit of detections was 0.79 ng mL-1, and 0.86 ng mL-1, respectively. The reproducibility of the AFP immunosensor was tested for 50 ng mL-1 AFP concentration and relative standard deviation (R.S.D.) values were calculated as 4.06% (n = 10) and 3.68% (n = 10) for DPV and SWV methods, respectively. The operation stability of the developed AFP immunosensor was examined by voltammetric methods for 60 days, and the storage stability for 12 weeks. The prepared AFP immunosensors were used for the analysis of different concentrations of AFP added to the human serum samples in a known amount and over 95% recoveries were obtained.
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Primary Language tr
Subjects Engineering, Chemical
Published Date Haziran-2021
Journal Section Kimya / Chemistry, Kimya Mühendisliği / Chemical Engineering
Authors

Orcid: 0000-0002-9146-0226
Author: Kadir ERDEM
Institution: ÇANKIRI KARATEKİN ÜNİVERSİTESİ, FEN FAKÜLTESİ
Country: Turkey


Orcid: 0000-0002-3381-7522
Author: Melike BİLGİ KAMAÇ (Primary Author)
Institution: Çankırı Karatekin Üniversitesi
Country: Turkey


Supporting Institution ÇANKIRI KARATEKİN ÜNİVERSİTESİ
Project Number FF61218209
Thanks Bu çalışma, Çankırı Karatekin Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından “FF61218209” kodlu proje ile desteklenmiştir.
Dates

Application Date : September 16, 2020
Acceptance Date : January 11, 2021
Publication Date : June 1, 2021

Bibtex @research article { jist795639, journal = {Journal of the Institute of Science and Technology}, issn = {2146-0574}, eissn = {2536-4618}, address = {}, publisher = {Igdir University}, year = {2021}, volume = {11}, pages = {1279 - 1292}, doi = {10.21597/jist.795639}, title = {Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi}, key = {cite}, author = {Erdem, Kadir and Bilgi Kamaç, Melike} }
APA Erdem, K , Bilgi Kamaç, M . (2021). Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi . Journal of the Institute of Science and Technology , 11 (2) , 1279-1292 . DOI: 10.21597/jist.795639
MLA Erdem, K , Bilgi Kamaç, M . "Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi" . Journal of the Institute of Science and Technology 11 (2021 ): 1279-1292 <https://dergipark.org.tr/en/pub/jist/issue/61423/795639>
Chicago Erdem, K , Bilgi Kamaç, M . "Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi". Journal of the Institute of Science and Technology 11 (2021 ): 1279-1292
RIS TY - JOUR T1 - Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi AU - Kadir Erdem , Melike Bilgi Kamaç Y1 - 2021 PY - 2021 N1 - doi: 10.21597/jist.795639 DO - 10.21597/jist.795639 T2 - Journal of the Institute of Science and Technology JF - Journal JO - JOR SP - 1279 EP - 1292 VL - 11 IS - 2 SN - 2146-0574-2536-4618 M3 - doi: 10.21597/jist.795639 UR - https://doi.org/10.21597/jist.795639 Y2 - 2021 ER -
EndNote %0 Iğdır Üniversitesi Fen Bilimleri Enstitüsü Dergisi Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi %A Kadir Erdem , Melike Bilgi Kamaç %T Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi %D 2021 %J Journal of the Institute of Science and Technology %P 2146-0574-2536-4618 %V 11 %N 2 %R doi: 10.21597/jist.795639 %U 10.21597/jist.795639
ISNAD Erdem, Kadir , Bilgi Kamaç, Melike . "Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi". Journal of the Institute of Science and Technology 11 / 2 (June 2021): 1279-1292 . https://doi.org/10.21597/jist.795639
AMA Erdem K , Bilgi Kamaç M . Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi. Iğdır Üniv. Fen Bil Enst. Der.. 2021; 11(2): 1279-1292.
Vancouver Erdem K , Bilgi Kamaç M . Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi. Journal of the Institute of Science and Technology. 2021; 11(2): 1279-1292.
IEEE K. Erdem and M. Bilgi Kamaç , "Kanser Biyobelirteci Alfa-Fetoproteinin Elektrokimyasal Tayini İçin Tek Kullanımlık Etiketsiz Yeni AFP İmmünosensörünün Geliştirilmesi", Journal of the Institute of Science and Technology, vol. 11, no. 2, pp. 1279-1292, Jun. 2021, doi:10.21597/jist.795639