Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

A Non-invasive Detection Technique of Adenocarcinoma with the Use of Streptavidin-Coated Iron Nanoparticles and Biotinylated-Antibodies and the Primary-Secondary Approach

Version 1 : Received: 22 June 2021 / Approved: 23 June 2021 / Online: 23 June 2021 (09:17:59 CEST)

How to cite: Venkatraman, A. A Non-invasive Detection Technique of Adenocarcinoma with the Use of Streptavidin-Coated Iron Nanoparticles and Biotinylated-Antibodies and the Primary-Secondary Approach. Preprints 2021, 2021060553 (doi: 10.20944/preprints202106.0553.v1). Venkatraman, A. A Non-invasive Detection Technique of Adenocarcinoma with the Use of Streptavidin-Coated Iron Nanoparticles and Biotinylated-Antibodies and the Primary-Secondary Approach. Preprints 2021, 2021060553 (doi: 10.20944/preprints202106.0553.v1).

Abstract

The streptavidin and biotin interaction is one of the strongest non-covalent interactions in nature. As a result, this non-covalent interaction has been of great interest when it comes to biochemical assays, diagnosis of diseases, and cell-targeted drug delivery. Past research has proven that biotin-streptavidin is useful in biosensor development to improve the detection of a system when conjugated to nanoparticles. This study aims to prove that streptavidin-coated nanoparticles can be conjugated with biotinylated antibodies using the primary-secondary method to non-invasively detect adenocarcinoma in-vitro. While the use of nanoparticles is not uncommon to the diagnostics area of scientific research, the technique this research aims to investigate is a non-invasive one, utilizing the primary-secondary method. Specifically, the increased stability of fluorophores when bound to antibodies as opposed to nanoparticles directly can be indicative of the particles conjugated through the primary-secondary method’s ability to specifically bind to overexpressed transferrin receptors in the A549 cell line. In this paper, streptavidin-coated nanoparticles were conjugated with biotinylated anti-transferrin receptor antibodies and AlexaFluor-488 secondary antibodies were used to enable fluorescence-based detection. The efficiency of these particles were observed quantitatively through a plate reader and qualitatively through a fluorescence microscope. I demonstrated that these nanoparticles are able to specifically bind to the target proteins in this study. These findings contribute to the field of nanoparticle diagnostics and can be extended to different diseases caused by overexpression of proteins in the future. While this was conducted in-vitro, conjugates can be prepared to detect cancer in-vivo and can be tested with magnetic relaxometry in the future.

Subject Areas

Iron-oxide nanoparticles, primary-secondary method, diagnostics of cancer, A549 cell line, transferrin protein receptors, adenocarcinoma, biotin-streptavidin, biotinylated antibodies

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