Characterization of Sm14 related components in different helminths by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting analysis

Thaumaturgo, Nilton; Vilar, Mônica Magno; Edelenyi, Ricardo; Tendler, Míriam

doi:10.1590/s0074-02762002000900024

Cited by 9 publications

(4 citation statements)

References 19 publications

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“…Ammonium sulphate precipitation (Nowotny, 1979 ) and caprylic acid purification (Nowotny, 1979 ) were used to purify the animal sera (Mckinney and Parkinson, 1987 ). Purified antibodies were tested for protein content (Bradford, 1976 ), and molecular weight was calculated using a 12.5 percent SDS-PAGE under reducing conditions (Myers, 1995 ; Thaumaturgo et al 2002 ). Indirect ELISA was used to test the reactivity of pure IgG pAbs.…”

Section: Methodsmentioning

confidence: 99%

Role of anti-Giardia recombinant cyst wall protein IgG polyclonal antibodies in diagnosis and protection

et al. 2022

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Giardia duodenalis (G. duodenalis) is an infectious protozoan that has a global distribution especially in the hot climate. Around 200 million people are infected worldwide annually by Giardia, but infection is not always accompanied by symptoms, especially in endemic countries. Using traditional microscopy techniques in diagnosis, both in stool and water samples were less sensitive when compared to immunological methods; and the need for new diagnostic methods was necessary. Also, protection from infection is required in endemic areas. Therefore, the study aimed to produce anti- G. duodenalis IgG polyclonal antibodies (pAbs) by immunizing rabbit by G. duodenalis cyst recombinant protein. The produced antibodies were evaluated in the detection of G. duodenalis antigens in patients’ stool and water samples from endemic areas across River Nile; where pAbs were used as a coating and a peroxidase conjugate antibody in sandwich ELISA. Moreover, pAbs were tested for the protection of mice from giardiasis. Sandwich ELISA using pAb has succeeded in the detection of G. duodenalis coproantigens in stool samples by a sensitivity of 97% and a specificity of 92.72%. Moreover, G. duodenalis cyst was detected in only seven water samples by ordinary microscopy; while sandwich ELISA revealed nineteen positive results. IgG pAb (1/200 µg/ml) protected mice from giardiasis; which was evident from the reduction in cysts and trophozoites numbers. We recommended the use of sandwich ELISA to monitor water quality, investigate environmental contamination and diagnosis in patients' stools. The pAbs can be prepared in large amount and used in field diagnosis and protection. This will help in the early diagnosis of G. duodenalis in water, which in turn can control outbreaks in rural areas.

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Section: Methodsmentioning

confidence: 99%

Role of anti-Giardia recombinant cyst wall protein IgG polyclonal antibodies in diagnosis and protection

et al. 2022

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“…The protein content of the prepared Ag was described by the method of Bradford (1976). Ion exchange chromatography (Sheehan and Gerald 1996), gel filtration chromatography on a Sephacryl S-200 HR column, and sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (Thaumaturgo et al 2002) were all used to purify CP. Indirect ELISA was utilized to test the reactivity of CP Ags (Engvall and Perlman 1971).…”

Section: S Mansoni Cp Antigen Preparationmentioning

confidence: 99%

Preparation of polyclonal anti-Schistosoma mansoni cysteine protease antibodies for early diagnosis

Farid

2023

Appl Microbiol Biotechnol

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In many parts of the tropics, schistosomiasis is a major parasitic disease second only to malaria as a cause of morbidity and mortality. Diagnostic approaches include microscopic sampling of excreta such as the Kato-Katz method, radiography, and serology. Due to their vital role in many stages of the parasitic life cycle, proteases have been under investigation as targets of immunological or chemotherapeutic anti-Schistosoma agents. Five major classes of protease have been identified on the basis of the peptide hydrolysis mechanism: serine, cysteine, aspartic, threonine, and metalloproteases. Proteases of all five catalytic classes have been identified from S. mansoni through proteomic or genetic analysis. The study aimed to produce polyclonal antibodies (pAbs) against schistosomal cysteine proteases (CP) to be used in the diagnosis of schistosomiasis. This study was conducted on S. mansoni-infected patients from highly endemic areas and from outpatients’ clinic and hospitals and other patients infected with other parasites (Fasciola, hookworm, hydatid, and trichostrongyloids). In this study, the produced polyclonal antibodies against S. mansoni cysteine protease antigens were labeled with horseradish peroxidase (HRP) conjugate and used to detect CP antigens in stool and serum samples of S. mansoni-infected patients by sandwich ELISA. The study involved 200 S. mansoni-infected patients (diagnosed by finding characteristic eggs in the collected stool samples), 100 patients infected with other parasites (Fasciola, hookworm, hydatid, and trichostrongyloids), and 100 individuals who served as parasite-free healthy negative control. The prepared pAb succeeded in detecting CP antigens in stool and serum samples of S. mansoni-infected patients by sandwich ELISA with a sensitivity of 98.5% and 98.0% respectively. A positive correlation was observed between S. mansoni egg counts and both stool and serum antigen concentrations. Purified 27.5 kDa CP could be introduced as a suitable candidate antigen for early immunodiagnosis using sandwich ELISA for antigen detection. Key points • Detection of cysteine protease antigens can replace parasitological examination. • Sandwich ELISA has a higher sensitivity than microscopic examination of eggs. • Identification of antigens is important for the goal of obtaining diagnostic tools.

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“…Cytoplasmic FABPs allow schistosomes to acquire fatty acids and cholesterol from the host due to a lack of schistosome oxygen-dependent synthetic pathways essential for membrane formation, protein anchoring, maturation, and egg production (75)(76)(77)(78)(79). Sm FABP shares 91%, 45%, 39% and 49% sequence homology with FABPs of Sj, Echinococcus granulosus, Fasciola hepatica, and Clonorchis sinensis, respectively; phylogenetic proximity may suggest potential for cross-species protection (80)(81)(82)(83).…”

Section: S Mansoni Fatty Acid-binding Protein: Sm14mentioning

confidence: 99%

Recent Advances and Methodological Considerations on Vaccine Candidates for Human Schistosomiasis

et al. 2021

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Schistosomiasis remains a neglected tropical disease of major public health concern with high levels of morbidity in various parts of the world. Although considerable efforts in implementing mass drug administration programs utilizing praziquantel have been deployed, schistosomiasis is still not contained. A vaccine may therefore be an essential part of multifaceted prevention control efforts. In the 1990s, a joint United Nations committee promoting parasite vaccines shortlisted promising candidates including for schistosomiasis discussed below. After examining the complexity of immune responses in human hosts infected with schistosomes, we review and discuss the antigen design and preclinical and clinical development of the four leading vaccine candidates: Sm-TSP-2 in Phase 1b/2b, Sm14 in Phase 2a/2b, Sm-p80 in Phase 1 preparation, and Sh28GST in Phase 3. Our assessment of currently leading vaccine candidates revealed some methodological issues that preclude a fair comparison between candidates and the rationale to advance in clinical development. These include (1) variability in animal models - in particular non-human primate studies - and predictive values of each for protection in humans; (2) lack of consensus on the assessment of parasitological and immunological parameters; (3) absence of reliable surrogate markers of protection; (4) lack of well-designed parasitological and immunological natural history studies in the context of mass drug administration with praziquantel. The controlled human infection model - while promising and unique - requires validation against efficacy outcomes in endemic settings. Further research is also needed on the impact of advanced adjuvants targeting specific parts of the innate immune system that may induce potent, protective and durable immune responses with the ultimate goal of achieving meaningful worm reduction.

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Characterization of Sm14 related components in different helminths by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting analysis

Cited by 9 publications

References 19 publications

Role of anti-Giardia recombinant cyst wall protein IgG polyclonal antibodies in diagnosis and protection

Role of anti-Giardia recombinant cyst wall protein IgG polyclonal antibodies in diagnosis and protection

Preparation of polyclonal anti-Schistosoma mansoni cysteine protease antibodies for early diagnosis

Recent Advances and Methodological Considerations on Vaccine Candidates for Human Schistosomiasis

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