Development of Sars-Cov-2 Antigen Detection Kit Based on Immunoglobulin Y (Igy) Using Surface Plasmon Resonance (Spr)
Thyazen Alhakimi1,6*
, Toto Subroto2,3, Muhammad Yusuf2,3, Wyanda Arnafia5, Ani Melani Maskoen4 , Suwendar4,Gilang Gumilar7,8 and Isa Anshori91Biotechnology Program, Postgraduate School, Universitas Padjadjaran, Jl. Dipati Ukur 35, Bandung 40132, Indonesia
2Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Sumedang 45363, Indonesia
3Research Center of Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, Bandung 40132, Indonesia
4Faculty of Medicine, Universitas Padjadjaran, Sumedang 45363, Indonesia
5Research and Development Division, Tekad Mandiri Citra Inc., Bandung 40292, Indonesia
6Department of pharmacy, Faculty of Mathematics and Natural Sciences, Universitas Islam Bandung, Bandung 40116, Indonesia
7Welding and Fabrication Engineering Technology Department, Institut Teknologi Sains Bandung, Bekasi 17530, Indonesia
8Advanced Functional Materials (AFM) Laboratory, Engineering Physics Department, Institut Teknologi Bandung, Bandung 40132, Indonesia
9Lab-on-Chip Group, Biomedical Engineering Department, Bandung Institute of Technology, Bandung 40312, Indonesia
Corresponding Author E-mail: thyazen16001@mail.unpad.ac.id
Abstract: SARS disease reappeared at the end of 2019 with a new name as Coronavirus Disease 2019 (COVID-19) caused by a new virus called SARS-CoV-2. This virus has spread throughout the world until recently and caused massive deaths and losses. The nucleic acid test in the form of real-time reverse transcriptase-polymerase chain reaction (RT-PCR) is very important to diagnose COVID-19 in patients, but this method has several drawbacks such as operators who have to be trained, the diagnosis results appear in a relatively long time, and the examination price relatively expensive. This research was conducted to produce immunoglobulin Y (IgY) extracted from chicken egg yolk targeting the S-protein receptor-binding domain (RBD) on SARS-CoV-2 as a component of the surface plasmon resonance (SPR) SARS-CoV-2 antigen detection kit. This research was started by extracting IgY from hyperimmune chicken egg yolks with the polyethylene glycol (PEG) precipitation method and continued with dialysis. The extracted IgY was further purified using thiophilic adsorption chromatography and concentrated by using Amicon® Ultra-15 ultrafiltration. The IgY activity against SARS-CoV-2 RBD was tested qualitatively using the agar gel precipitation test (AGPT) technique and the total protein content was determined using the Lowry method. IgY was tested for its affinity against SARS-CoV-2 RBD using SPR. The IgY concentration obtained was 11 mg/mL. The AGPT result showed the presence of IgY activity against SARS-CoV-2 RBD isolated from egg yolk and chicken serum after 8 weeks after the first vaccination of chickens. The SDS-PAGE results showed a very clear band of IgY characters. The obtained IgY showed adequate interaction with commercial SARS-CoV-2 RBD on an SPR device. The purified IgY was able to bind with protein-S RBD and showed a fairly good affinity for the SARS-CoV-2 antigen sample. The results of these observations indicate that IgY anti-S-protein SARS-CoV-2 can be produced and purified from chicken egg yolk and can be used as a diagnostic component to detect SARS-CoV-2 antigen, especially on SPR.
Keywords: Antibody (IgY); Covid-19; SARS-CoV-2; IgY anti- SARS-CoV-2; Surface Plasmon Resonance (SPR); SARS-CoV-2 RBD Back to TOC