Recombinant antibody phage display technology is a vital tool that facilitates identification of specific binding molecules to a target enabling the rapid generation and selection of high affinity, fully human, or mouse antibody product candidates essentially directed towards disease target appropriate for antibody therapy. In this study, a recombinant single-chain Fv antibody fragment (scFv) A11 was isolated from immune spleen cells obtained from mice immunized with inactivated rabies virus (Pasteur strain) using standard methodology and was characterized for its specificity towards the rabies virus glycoprotein. Epitope mapping using peptide libraries and truncated glycoprotein polypeptides suggested that A11 bound to the antigenic site II of rabies glycoprotein against which a majority of rabies virus neutralizing antibodies are directed. The use of the above technology could, therefore, allow development of scFvs with different specificities against the rabies glycoprotein as an alternative to the more cumbersome protocols used for the development of monoclonal antibodies. 1. Introduction Rabies is a viral, zoonotic, and invariably fatal neuroinvasive disease of humans caused by the bite of rabid animal. More than 55,000 deaths occur annually worldwide in spite of the use of postexposure therapy preventive measures, making rabies one of the major causes of human mortality despite significant scientific progress [1, 2]. Administration of antivirus immunoglobulins includes the use of both human and equine antirabies immunoglobulins along with vaccination and is the only strategy recommended by WHO for the postexposure prophylactic treatment of rabies [3]. Most of these immunoglobulins are plasma-derived, polyclonal products obtained from human and animal donors vaccinated against rabies which can be produced in limited amounts and suffer from potential drawbacks such as batch-to-batch variation and the risk of contamination from other pathogens [4, 5]. Several conventional monoclonal antibodies (MAbs) have been generated against different viruses [6] and their utilization tends to be limited in clinical applications because of possible viral contamination and high cost involved in MAb preparations. To overcome these problems, generation of single chain antibody fragments (scFv) through phage display technology has been utilized as the methodology of choice [7–9]. Use of phage display technology as a powerful in vitro tool for production of therapeutically important antibodies against viral pathogens on the surface of bacteriophage provides an
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