Brenda Song, Pei Chui
(2024)
Development And Characterization Of Monoclonal Antibodies Against
Ancylostoma Caninum Ancylostoma-Secreted Protein 5 (Asp5) By Phage Display.
Masters thesis, Universiti Sains Malaysia.
Abstract
Parasitic nematodes such as Ancylostoma caninum engages in a complex life cycle, in which it can impact the hosts at its various stages of development and infection. During infective stage, Ancylostoma caninum releases a multitude of proteins to modulate the host’s immune response, which ensuring its survival within the host environment. Among these proteins, Ancylostoma-secreted protein 5 (ASP5) is critically involved in the parasite-host interaction such as the immunomodulation, tissue invasion and facilitating the blood feeding process, hence making it a promising target for intervention strategies aimed at controlling hookworm infections among canines. By harnessing phage display technology, this study aims to develop monoclonal antibodies targeting Ancylostoma-secreted protein 5 (ASP5), by employing the human naïve scFv antibody library through in vitro biopanning. The library was subjected to a total of three rounds of panning against 10 μg of ASP5 antigen, with progressively stringent washing conditions (10, 20 and 30 washes respectively) to isolate the clones. This study delves into Ancylostoma-secreted protein 5 as an antigenic candidate for monoclonal antibody development in which it was expressed using the bacterial strain of BL21(DE3) and purified with Immobilized Metal Affinity Chromatography (IMAC) purification method using the nickel-charged affinity resin. The developed antibody was characterized on its antibody-antigen interactions, including cross-reactivity and binding strength via Enzyme-linked immunosorbent assay (ELISA).
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