Construction of cloudy catshark (Scyliorhinus torazame) variable new antigen receptor (vNAR) library using phage display and application for antigen detection

Abstract

Monoclonal antibodies (mAbs) have been widely applied as diagnostic and therapeutic platforms. However, these conventional mAbs have shown several shortcomings, such as their large size and difficult to store. To overcome the problems of the mAbs, second-generation antibodies based on the single-chain fragment variable (scFv) have been developed but, unstability and lacking avidity are the shortcomings of scFV antibodies. Nowadays, the Immunoglobulin New Antigen Receptor (IgNAR) founded in sharks is considered as an attractive candidate for applicable antibody technologies because of several advantageous properties such as cryptic antigen recognition domain structure, small molecular size, fast secretion, stability. Phage display is a rapid, cost-effective technology to display the antibody, so we constructed the phage library display using the vNAR gene from cloudy catshark (Scyliorhinus torazame) with the phage display platform. Furthermore, we performed screening about Lysozyme A to check the operation of constructed phage library and did another screening about VHSV to confirm the applicability of specific pathogens. Based on the results of screening about Lysozyme A and VHSV, we confirmed that the generated phage library display was working well and checked the applicability to a specific antigen. However, a present study about the VHSV is incomplete so, it is necessary to find more VHSV positive clones through the ELISA assay, and further studies using VHSV positive clones such as neutralization assay, diagnostic tests about VHSV infected cells and tissues. Consequently, it is necessary to perform further studies, but the constructed phage library display can be applied to specific antigens such as other Rhabdovirus, therapeutic and diagnostic, and will enable rapid, cost-effective production of antibodies with specificity.