Abstract
Helminth parasite infections are significantly impacting global health, with more than two billion infections worldwide with a high morbidity rate. The complex life cycle of the nematodes has made host immune response studies against these parasites extremely difficult. In this study, we utilized two phage antibody libraries; the immune and naïve library were used to identify single chain fragment variable (scFv) clones against a specific filarial antigen (BmR1). The V-gene analysis of isolated scFv clones will help shed light on preferential VDJ gene segment usage against the filarial BmR1 antigen in healthy and infected states. The immune library showed the usage of both lambda and kappa light chains. However, the naïve library showed preferential use of the lambda family with different amino acid distributions. The binding characteristics of the scFv clones identified from this work were analyzed by immunoassay and immunoaffinity pull down of BmR1. The work highlights the antibody gene usage pattern of a naïve and immune antibody library against the same antigen as well as the robust nature of the enriched antibodies for downstream applications.
Highlights
Human lymphatic filariasis infection remains a major public health problem, mainly in tropical countries
The World Health Organization (WHO) has identified lymphatic filariasis as the second leading cause of permanent and long-term disability in the world, after leprosy. It is caused by microscopic, thread-like worms, namely Wuchereria bancrofti (W. bancrofti), Brugia malayi (B. malayi), and Brugia timori (B. timori)
Phage particles displaying individual human single chain fragment variable (scFv) antibody fragments were selected by three rounds of biopanning on Maxisorp plates coated with BmR1 antigen using both naïve and immune libraries
Summary
Human lymphatic filariasis infection remains a major public health problem, mainly in tropical countries. In order to study the different antibody gene usage patterns of a healthy and parasite-infected population, the IgM and IgG repertoires would be ideal for use. The IgG and IgM repertoire has to be replicated in vitro in order to investigate the differences experimentally This can be achieved by using phage display technology where the repertoire is captured in a collection of cloned antibody sequences as a fusion to phage coat proteins to produce a diverse antibody library for panning [6]. An immune library for lymphatic filariasis (IgG isotype) [13] and a naïve library (IgM isotype) [14] was used to compare the ability of each library repertoire to generate monoclonal scFv clones against a specific filarial target. The antibodies developed have potential diagnostic applications, evident from the results of the binding studies
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