RESUMO
Echinococcus granulosus is a worldwide zoonotic infection that causes human cystic echinococcosis (CE) or hydatid disease. The present study describes the isolation and production of a monoclonal antibody against recombinant AgB protein using the developed Human AntibodY Disease ENhanced (HAYDEN)-Filariasis library. The DNA sequences of the isolated clones were analyzed, followed by gene analysis and binding assays. Clone E1 showed a full-length sequence and represents the IgHV5-LV3 antibody gene family. The antibody protein yield was satisfactory, and it reacted specifically against rAgB. The novel E1 protein is potentially useful for the development of an antigen detection assay for CE. The ability of the Brugia malayi immune antibody library to isolate antibodies against Echinococcus granulosus antigens highlights the broad coverage of immune antibody libraries.
Assuntos
Anticorpos Anti-Helmínticos/imunologia , Anticorpos Monoclonais/isolamento & purificação , Brugia Malayi/imunologia , Echinococcus granulosus/imunologia , Lipoproteínas/imunologia , Animais , Anticorpos Monoclonais/imunologia , Sequência de Bases , Western Blotting , Brugia Malayi/genética , Equinococose/diagnóstico , Echinococcus granulosus/genética , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Epitopos/imunologia , Humanos , Lipoproteínas/genéticaRESUMO
Toxocariasis is a widespread zoonotic parasitic disease with a significant socioeconomic impact, particularly on underprivileged communities. Limitations of existing diagnostic tools and vague presenting symptoms may lead to misdiagnosis, thus underestimating the actual global impact of the disease. The present study describes the isolation and production of novel recombinant monoclonal antibodies against Toxocara canis recombinant TES-26 antigen (rTES-26) utilizing a human helminth scFv phage display library. The isolated antibody clones were characterized based on their gene sequences and binding characteristics. Three clones representing unique gene families (clone 48: IgHV3-LV1; clone 49: IgHV3-LV3; clone 50: IgHV6-LV3) were isolated, but only clones 48 and 49 showed successful insertion of the full-length scFv antibody sequence after sub-cloning. Both clones produced antibody proteins of good solubility and satisfactory yield and purity. Binding assays via Western blot and ELISA using rTES-26 and Toxocara canis native protein showed that both monoclonal antibodies were highly specific and sensitive to the target antigen. A preliminary antigen detection ELISA showed the diagnostic potential of the monoclonal antibody proteins. The proteins can also be useful in studying host−parasite interactions and therapeutic applications.
RESUMO
Antibodies have different chemical properties capable of targeting a diverse nature of antigens. Traditionally, immune antibody libraries are perceived to be disease-specific with a skewed repertoire. The complexity during the generation of a combinatorial antibody library allows for a skewed but diverse repertoire to be generated. Strongyloides stercoralis is a parasite that causes strongyloidiasis, a potentially life-threatening disease with a complex diagnosis that impedes effective control and treatment of the disease. This study describes the isolation of monoclonal antibodies against S. stercoralis NIE recombinant protein using an immune antibody phage display library derived from lymphatic filaria-infected individuals. The isolated antibody clones showed both lambda and kappa light chains gene usage, with diverse amino acid distributions. Structural analysis showed that electropositivity and the interface area could determine the binding affinity of the clones with NIE. The successful identification of S. stercoralis antibodies from the filarial immune library highlights the breadth of antibody gene diversification in an immune antibody library that can be applied for closely related infections.
Assuntos
Anticorpos Anti-Helmínticos/imunologia , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos/imunologia , Biblioteca de Peptídeos , Anticorpos de Cadeia Única/imunologia , Strongyloides/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Anti-Helmínticos/química , Anticorpos Anti-Helmínticos/genética , Anticorpos Anti-Helmínticos/isolamento & purificação , Anticorpos Monoclonais/química , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/isolamento & purificação , Antígenos de Helmintos/química , Antígenos de Helmintos/imunologia , Sítios de Ligação , Técnicas de Visualização da Superfície Celular , Modelos Moleculares , Ligação Proteica , Conformação Proteica , Proteínas Recombinantes/genética , Análise de Sequência de DNA , Anticorpos de Cadeia Única/química , Anticorpos de Cadeia Única/genética , Anticorpos de Cadeia Única/isolamento & purificação , Relação Estrutura-AtividadeRESUMO
Hydatid disease is not endemic in Malaysia; however, its migrant workers originate from neighboring countries where the disease is prevalent. Thus, this study was aimed at investigating the seroprevalence of hydatid disease among the workers. A total of 479 migrant workers were screened for hydatid disease. The sociodemographic information was collected, and serum samples were tested with a rapid dipstick test for hydatid disease called Hyd Rapid™. The present study showed that 13.6% of the migrant workers were found to be seropositive for hydatid disease. The highest seroprevalence was seen among Indian workers (29.41%), followed by Myanmarese (21.43%), Bangladeshis (14.92%), Nepalese (10.68%), and Indonesian (10.66%). This is the first study that highlights the likely presence of hydatid disease among the migrant workers in Malaysia, which may be of interest to the health authorities.