CONSTRUCTION OF LATERAL FLOW IMMUNOASSAY BASED ON IMMUNOGLOBULIN Y ANTI-SARS-CoV-2 AS A DIAGNOSTIC KIT TO DETECT CORONAVIRUS DISEASE-19 (COVID-19)

Abstract

Coronavirus disease 2019 (COVID-19) is a disease caused by a highly contagious viral infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 was first reported in Wuhan, China then spread throughout the world until today causing massive deaths and losses. In addition to the development of drugs carried out, it is also necessary to design a diagnostic kit to diagnose this disease to control its spread. Several diagnostic methods have been devised to diagnose COVID-19. Lateral flow immunoassay (LFIA) is a diagnostic method that meets the ASSURED criteria (Affordable, Sensitive, Specific, User-friendly, Rapid and robust, Equipment-free, Deliverable to end-users) as an effective and efficient diagnostic kit. One of the most important parts of the LFIA is the test line and conjugate pad which have a direct impact on LFIA sensitivity. The main component of the test line is the antibody and of the conjugate pad is the gold nanoparticle-antibody conjugate. This research was conducted to develop LFIA to diagnose COVID-19 based on IgY anti-SARS-CoV-2 as an antibody component because IgY production from chickens is more efficient than mAb IgG production using hybridoma techniques from mammals. The research was carried out by purifying IgY from immunized chicken egg yolk with PEG6000 extraction and thiophilic adsorption chromatography, characterization its molecular weight profile with SDS-PAGE and its affinity with Surface Plasmon Resonance (SPR), synthesis of gold nanoparticles (AuNP), AuNP-IgY conjugation, IgY immobilization against nitrocellulose membranes and AuNP-IgY conjugates against the conjugate pad, LFIA construction, and LFIA testing against SARS-CoV-2 samples. The result shows that the obtained IgY purity was 91% which had a good affinity against SARS-CoV-2 according to the SPR sensorgram. The constructed LFIA was succeed to detect the SARS-CoV-2. These methods can be used to develop LFIA based on IgY anti-SARS-CoV-2 for diagnosing COVID-19