Abstract
<p><span lang="EN-US">The need for rapid diagnostic tools or point- of- care diagnostic tests for Avian Influenza in Indonesia is very high and the price of these imported diagnostic tools is very expensive. As a result, a large budget requires to provide the needs. The main component of a rapid diagnostic tool is the monoclonal antibody (mAb) specifically recognized influenza viruses. The objective of this study was to produce mAb that can recognize all subtypes of Avian Influenza viruses using the phage display technology. Influenza-A focused scFv commercial library was panned using alternating recombinant H1N1 NP and H5N1 virions. Whereas, bacteriophages bound to the panning baits were eluted with serum from H5N1-infected chickens. Phagemid from suppressor E. coli (TG1) infected with bacteriophage displaying anti-NP on its surface was isolated and then transformed into a non-suppressor E. coli (HB2151) to express NP-scFv. Monoclonal NP-scFv antibody with a molecular weight of about 27 kDa was purified from the culture supernatant using a nickel-chromatography column. The amount of pure NP-scFv obtained was around 1.2 mg /L culture. As an additional component for its use in immunoassays, antibody to NP-scFv was produced in rabbits. The generating polyclonal antibody recognized the NP-scFv specifically and sensitively. The anti-NP-scFv monoclonal antibody and the anti rabbit scFv polyclonal antibody produced in this study are envisaged appropriate for the development of diagnostic tools for point-of-care for Avian Influenza.</span></p>
Highlights
The H5N1 avian influenza seems to be one of the most devastating zoonotic diseases ever known to date (FAO 2013)
Diagnostic tools for Influenza both for human and poultry are based on monoclonal antibody specific against the nucleoprotein of the type A Influenza virus (Tarigan 2016)
Expression of anti-nucleoprotein H1N1 Influenza virus (NP) scFv antibody in the present study revealed that the amount of scFv recovered from culture supernatant was larger than that from periplasmic compartment
Summary
The H5N1 avian influenza seems to be one of the most devastating zoonotic diseases ever known to date (FAO 2013). One of the main factors causing the rapid, wide spread of the disease was the delay in diagnosis and implementing actions to eradicate the disease. Diagnosis of infectious diseases that spread rapidly such as AI H5N1 requires the availability of rapid or point-of -care diagnostic (POC) tools. Most of the POC diagnostic tools for Influenza both for human and poultry are based on monoclonal antibody specific against the nucleoprotein of the type A Influenza virus (Tarigan 2016). The main advantages of using monoclonal include batch-to-batch homogeneity and excellent specificity. Polyclonal antibodies are much easier, cheaper and faster to produce but the variability between different batches produced in different animals at different times is unavoidable. Since polyclonal antibody comprises huge number of JITV Vol 24 No 1 Th. 2019: 29-38 antibodies recognizing different epitopes, cross-reaction is inescapable (Liu 2014; Shalit et al 1985)
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