DEVELOPMENT OF POLYCLONAL IMMUNOGLOBULIN Y (IGY) ANTIBODIES TARGETING SARS-COV-2 MULTI-EPITOPE ANTIGEN FROM CHICKEN EGG YOLK

• Main Author: Naisanu, Katerina
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/82762
• Institution: Institut Teknologi Bandung

Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2) is the primary cause of the Coronavirus disease-2019 (COVID-19) pandemic that affects the lives of millions of people around the world with high infectivity and mortality rates. One way to detect SARS-CoV-2 for initial testing is with a Rapid Diagnostic Test (RDT) that uses specific antibodies to bind the SARS-CoV-2 virus. Generally, IgG from mammals is used as a specific antibody to bind the target antigen in the RDT developed, but it is known to take a long time to develop at a high cost. Therefore, a more efficient and economical method is needed with the development of polyclonal IgY from chicken egg yolk. In addition, the use of antigens for immunization also affects the immunogenicity of antibodies so that in this study antigens designed from SARS-CoV-2 protein S and NSP3 were used, namely SARS-CoV-2 multi-epitope peptides that can induce the host humoral and cellular immune systems. Four chickens were immunized intramuscularly using the SARS-CoV-2 multi-epitope peptide antigen and one chicken was used as control, which was not immunized. IgY was isolated from chicken eggs and then further characterized using SDS-PAGE method. Antigenicity testing of IgY was carried out by ELISA method using SARS-CoV-2 multi-epitope peptide antigen and COVD-19 positive patient samples. The results showed that the suspected IgY protein bands were observed on SDS-PAGE gels with two different molecular weights, namely heavy chain (H) at a size of about 68 kDa and light chain (L) at a size of 25 kDa. ELISA test results showed that IgY anti multiepitope peptides can recognize SARS-CoV-2 multiepitope peptide antigens, when compared to control IgY (IgY from non-immunized chickens) (p<0.05). Meanwhile, the results of testing polyclonal IgY anti-multiepitope peptides with COVID-19 positive patient samples at various concentrations (1:100; 1:1000; 1:10,000) showed an average value with significance (p<0.05), but it is known that the developed polyclonal IgY anti-multiepitope peptides still need to be tested further ELISA system because the titer of the test results is still below the detection limit so further testing is needed on positive COVID-19 patient samples. Thus it can be concluded that the development of immunoglobulin Y (IgY) polyclonal antibodies against multiepitope peptides has been successfully developed, but further tests need to be carried out on the introduction of IgY polyclonal antibodies against SARS-CoV-2 multiepitope peptides to positive patient samples of COVID-19 to ensure its potential in the development of a Rapid Diagnostic Test (RDT).