Immunological tests for diarrhoea caused by diarrhoeagenic Escherichia coli targeting their main virulence factors

Piazza, Roxane M. F.; Caetano, Bruna Alves; Henrique, Camila; Luz, Daniela; Munhoz, Danielle Dias; Polatto, Juliana Moutinho; Rocha, Letícia B.; Silva, Miriam A.; Mitsunari, Thaís

doi:10.1016/bs.mim.2019.11.004

Cited by 6 publications

(3 citation statements)

References 85 publications

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“…albopictus yang telah melalui proses purifikasi kemudian dideteksi karakter imunogenesitasnya melalui analisis dot blot. Prinsip kerja dot blot ialah mendeteksi pengenalan molekul melalui ikatan antara antibodi dan antigen yang direaksikan diatas media membran (Piazza et al, 2020). Reaksi ikatan antara antigen dan antibodi divisualisasikan dengan kemunculan noktah gelap pada permukaan membran PVDF.…”

Section: Pembahasanunclassified

PURIFIKASI FRAKSI PROTEIN IMUNOGENIK 47 kDa DARI KELENJAR SALIVA Aedes albopictus SEBAGAI TARGET PENGEMBANGAN VAKSIN DENGUE BERBASIS VEKTOR

Wathon,

Rahmawati,

Oktarianti

et al. 2023

Berita Biologi

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Kapasitas vektorial Ae. albopictus telah diketahui sebagai vektor potensial virus dengue yang mengakibatkan DBD. Transmisi virus dengue terjadi ketika Ae. albopictus melakukan blood feeding ke manusia yang difasilitasi oleh aktivitas biologis protein pada kelenjar saliva vektor. Penelitian kami sebelumnya menunjukkan fraksi protein 47 kDa dari kelenjar saliva Ae. albopictus bersifat imunogenik. Protein 47 kDa merupakan serpins family yang diduga sebagai antikoagulan dan berperan sebagai protease inhibitor sehingga mempermudah transmisi virus dengue. Peran protein tersebut mengindikasikan adanya potensi dalam pengembangan vaksin penghambat transmisi patogen melalui vaksinasi melawan protein tersebut sehingga dapat menghambat transmisi dengue. Studi potensi protein tersebut memerlukan ekstrak murni, maka purifikasi protein target merupakan langkah yang sangat penting. Penelitian ini dimulai dengan landing collection dan rearing Ae. albopictus, isolasi dan ekstraksi protein kelenjar saliva, isolasi fraksi protein 47 kDa melalui SDS-PAGE, purifikasi protein, analisis dot blot dan western blot. Analisis SDS-PAGE menunjukkan pita tunggal protein target 47 kDa dari kelenjar saliva Ae. albopictus yang berhasil dipurifikasi menggunakan Electroeluter. Analisis dot blot menunjukkan protein 47 kDa terdeteksi bersifat imunogenik dan dikonfirmasi melalui analisis western blot bahwa protein target hasil purifikasi memiliki berat molekul 47 kDa.

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Section: Pembahasanunclassified

PURIFIKASI FRAKSI PROTEIN IMUNOGENIK 47 kDa DARI KELENJAR SALIVA Aedes albopictus SEBAGAI TARGET PENGEMBANGAN VAKSIN DENGUE BERBASIS VEKTOR

Wathon,

Rahmawati,

Oktarianti

et al. 2023

Berita Biologi

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“…One of the techniques in diagnostic kit manufacturing is based on dot-blotting immunoassay, which comprises four essential elements: holder, membrane, protein antigen, or any biological recognition elements, as well as buffer solution (Piazza et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

Suppression of Coffee-ring Effect (CRE) in the Development of Low-cost Diagnostic Kit

Ideris,

Abu Bakar,

Ahmad

et al. 2024

E-BPJ

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One of the applications of membrane technology is using the polymeric membrane as an adsorber or assay-capturing matrix in the diagnostic kits' assembly. This study explores the addition of NaCl into a protein solution to suppress the coffee-ring effect (CRE) in developing a low-cost diagnostic kit. The highest concentration of NaCl addition shows the optimum results with no formation of CRE and high color intensity (low grey scale value). Adding NaCl into the protein solution is a safe and cheap alternative for lowering the cost of assembly, benefiting people in low-resource places.

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“…A particularly important role is played by the immunoglobulin corona adsorbed on polymer microparticles referred to as immunolatexes [ 7 , 8 ]. Such conjugates find their application in a variety of tests, including those for bacterial infections such as E. coli and Salmonella [ 9 , 10 , 11 ], viral infections such as HIV [ 12 , 13 , 14 ] and more recently against SARS-CoV-2 [ 15 , 16 , 17 , 18 ]. The widespread application of these tests is primarily attributed to their simplicity and remarkably short execution time, typically taking just a few minutes [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

Mechanism of Anti-Salmonella Rabbit Immunoglobulin Adsorption on Polymer Particles

Żeliszewska,

Wasilewska,

Szych

et al. 2023

Biomolecules

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The adsorption of anti-Salmonella rabbit immunoglobulin (IgaR) on negatively charged polymer particles leading to the formation of immunolatex was studied using various techniques comprising atomic force microscopy (AFM) and laser Doppler velocimetry (LDV). Initially, the basic physicochemical properties of IgaR molecules and the particles, inter alia their electrophoretic mobilities, the zeta potentials and hydrodynamic diameters, were determined under different ionic strengths and pHs. Applying AFM, single immunoglobulin molecules adsorbed on mica were also imaged, which allowed to determine their size. The adsorption of the IgaR molecules on the particles leading to changes in their electrophoretic mobility was monitored in situ using the LDV method. The obtained results were interpreted applying a general electrokinetic model which yielded quantitative information about the molecule coverage on the particles. The obtained immunolatex was thoroughly characterized with respect to its acid–base properties and its stability upon storage. Notably, the developed procedure demonstrated better efficiency compared to commercially applied methods, characterized by a higher immunoglobulin consumption.

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Immunological tests for diarrhoea caused by diarrhoeagenic Escherichia coli targeting their main virulence factors

Cited by 6 publications

References 85 publications

PURIFIKASI FRAKSI PROTEIN IMUNOGENIK 47 kDa DARI KELENJAR SALIVA Aedes albopictus SEBAGAI TARGET PENGEMBANGAN VAKSIN DENGUE BERBASIS VEKTOR

PURIFIKASI FRAKSI PROTEIN IMUNOGENIK 47 kDa DARI KELENJAR SALIVA Aedes albopictus SEBAGAI TARGET PENGEMBANGAN VAKSIN DENGUE BERBASIS VEKTOR

Suppression of Coffee-ring Effect (CRE) in the Development of Low-cost Diagnostic Kit

Mechanism of Anti-Salmonella Rabbit Immunoglobulin Adsorption on Polymer Particles

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