1. Simple workflow to repurpose SARS-CoV-2 swab/serum samples for the isolation of cost-effective antibody/antigens for proteotyping applications and diagnosis
- Author
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Faezeh Ghorbanizamani, Simge Balaban Hanoglu, Kerem Tok, Duygu Harmanci, Ceren Durmus, Serap Evran, Hichem Moulahoum, Candan Çiçek, Rüçhan Sertöz, Bilgin Arda, Kutsal Turhan, Figen Zihnioglu, Suna Timur, and Tuncay Göksel
- Subjects
Coronavirus disease 2019 (COVID-19) ,Antigen/antibody purification ,Cost-Benefit Analysis ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Enzyme-Linked Immunosorbent Assay ,Lateral flow assay ,Calorimetry ,Antibodies, Viral ,Tandem mass spectrometry ,Biochemistry ,Antibodies ,Specimen Handling ,Workflow ,Analytical Chemistry ,Antigen ,Affinity chromatography ,Tandem Mass Spectrometry ,Information ,Humans ,Magnetic nanoparticle (MNP)–based separation ,Viremia ,Antigens, Viral ,Purification ,Cleavage ,Immunoassay ,Magnetic nanoparticle (MNP)-based separation ,antibody purification ,Mass spectrometry ,biology ,SARS-CoV-2 ,Chemistry ,COVID-19 ,Magnetic Nanoparticles ,Isolation (microbiology) ,Virology ,biology.protein ,Electrophoresis, Polyacrylamide Gel ,Antibody ,Research Paper - Abstract
Supply shortage for the development and production of preventive, therapeutic, and diagnosis tools during the COVID-19 pandemic is an important issue affecting the wealthy and poor nations alike. Antibodies and antigens are especially needed for the production of immunological-based testing tools such as point-of-care tests. Here, we propose a simple and quick magnetic nanoparticle (MNP)-based separation/isolation approach for the repurposing of infected human samples to produce specific antibodies and antigen cocktails. Initially, an antibody cocktail was purified from serums via precipitation and immunoaffinity chromatography. Purified antibodies were conjugated onto MNPs and used as an affinity matrix to separate antigens. The characterization process was performed by ELISA, SDS-PAGE, electrochemistry, isothermal titration calorimetry, and LC-Q-TOF-MS/MS analyses. The MNP-separated peptides can be used for mass spectrometry-based as well as paper-based lateral flow assay diagnostic. The exploitation of the current workflow for the development of efficient diagnostic tools, specific treatments, and fundamental research can significantly impact the present or eventual pandemic. This workflow can be considered as a two birds, one stone-like strategy., TUBITAK [120R021]; Republic of Turkey Ministry of Development [2010K120810/2020K12150700, 2016K121190], The study was supported by TUBITAK (Project number: 120R021). The Republic of Turkey Ministry of Development provided the support for the infrastructure of EGEMATAL (Ege University/Izmir) via 2010K120810/2020K12150700 and 2016K121190 grants.
- Published
- 2021