New highly sensitive sandwich ELISA system for soluble endoglin quantification in different biological fluids

Смирнов, И. В.; Gryazeva, Irina V.; Vasileva, Margarita Yurevna; Krutetskaia, Irina Yurevna; Shashkova, O. A.; Samoylovich, M. P.; Stolbovaya, A. Y.; Solodovnikova, Nellya Grigorevna; Zazerskaya, Irina E.; Sokolov, D. N.; Selkov, S. A.; Климович, В. Б.

doi:10.1080/00365513.2018.1516892

Cited by 12 publications

(11 citation statements)

References 33 publications

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“…To develop an effective sandwich ELISA, the pair of selected antibodies (capture and detection antibodies) is key in determining the sensitivity and specificity of the assay [33]. To date, traditional antibodies are universally used to develop sandwich ELISAs [8,9,34]. Yet, procedures of preparation, purification and labelling of traditional antibodies are complicated and costly, which is a heavy burden for the development of commercial sandwich ELISA kits [9,23,35].…”

Section: Discussionmentioning

confidence: 99%

“…To date, traditional antibodies are universally used to develop sandwich ELISAs [8,9,34]. Yet, procedures of preparation, purification and labelling of traditional antibodies are complicated and costly, which is a heavy burden for the development of commercial sandwich ELISA kits [9,23,35]. In addition, the double-antibody sandwich ELISA is not extensively used for clinically detection of antigens and diagnosis of disease due to its high cost [36].…”

Section: Discussionmentioning

confidence: 99%

“…In recent years, many commercial ELISA kits based on viral antigens and NDV-specific conventional antibodies have also been applied for the rapid diagnosis of NDV [22]. Yet, procedures of preparation, purification and reporter labelling of traditional antibodies are complicated and costly, which is a heavy burden for the development of commercial ELISA kits [9,23]. In this study, NDV specific nanobodies were obtained from a Bactrian camel immunized with inactivated NDV vaccines.…”

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confidence: 99%

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Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

Zhu

et al. 2020

Preprint

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Background: Traditional sandwich enzyme-linked immunosorbent assay (ELISA) using polyclonal and monoclonal antibodies as reagents presents several drawbacks, including limited amounts, difficulty in permanent storage, and required use of a secondary antibody. Nanobodies can be easily expressed with different systems and fused with several tags in their tertiary structure by recombinant technology, thus offering an effective detection method for diagnostic purposes. Recently, the fenobody (ferritin-fused nanobody) and RANbody (nanobody-fused reporter) have been designed and derived from the nanobody for developing the diagnostic immunoassays. However, there was no report about developing the sandwich ELISA using the fenobody and RANbody as pairing reagents.Results: A platform for developing a sandwich ELISA utilizing fenobody as the capture antibody and RANbody as the detection antibody was firstly designed in the study. Newcastle disease virus (NDV) was selected as the antigen, from which 13 NDV-specific nanobodies were screened from an immunized Bactrian camel. Then, 5 nanobodies were selected to produce fenobodies and RANbodies. The best pairing of fenobodies (NDV-fenobody-4, 800 ng/well) and RANbodies (NDV-RANbody-49, 1:10) was determined to develop the sandwich ELISA for detecting NDV. The detection limits of the assay were determined to be 22 of hemagglutination (HA) titers and 10 ng of purified NDV particles. Compared with two commercial assays, the developed assay shows higher sensitivity and specificity. Meanwhile, it exhibits 98.7% agreement with the HA test and can detect the reference NDV strains belonging to Class II but not Class I. Conclusions: In the presented study, the 13 anti-NDV nanobodies binding the NDV particles were first produced. Then, for the first time, the sandwich ELISA to detect the NDV in the different samples has been developed using the fenobody and RANbody as reagents derived from the nanobodies. Considering the rapidly increasing generation of nanobodies, the platform can reduce the cost of production for the sandwich ELISA and be universally used to develop assays for detecting other antigens.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

Zhu

et al. 2020

Preprint

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show abstract

“…of capture and reporter-labeled detection antigen-specific antibodies is essential and must be produced in the initial step [6,7]. While most sandwich ELISA kits and housed-methods employ conventional polyclonal and monoclonal antibodies as indispensable reagents, they present several drawbacks, including limited amounts, difficulty in permanent storage, and required use of a secondary antibody [8,9]. Hence, there is an urgent need to develop strategies for producing smaller size recombinant antibodies that are more easily produced, selected, and manipulated.…”

mentioning

confidence: 99%

“…Then, the sandwich ELISA for detecting H5N1 virus using the fenobody as capture antibody can be significantly improved the sensitivity. Yet, procedures of preparation, purification and reporter labelling of traditional antibodies are complicated and costly, which is a heavy burden for the development of commercial ELISA kits [9,25]. The RANbodies (nanobodies-fused reporter protein) [26,27] derived from nanobody can also overcome the drawback and there are no need to be reporter labelled the traditional antibodies.…”

mentioning

confidence: 99%

Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

Zhu

et al. 2020

J Nanobiotechnol

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Background: Traditional sandwich enzyme-linked immunosorbent assay (ELISA) using polyclonal and monoclonal antibodies as reagents presents several drawbacks, including limited amounts, difficulty in permanent storage, and required use of a secondary antibody. Nanobodies can be easily expressed with different systems and fused with several tags in their tertiary structure by recombinant technology, thus offering an effective detection method for diagnostic purposes. Recently, the fenobody (ferritin-fused nanobody) and RANbody (nanobody-fused reporter) have been designed and derived from the nanobody for developing the diagnostic immunoassays. However, there was no report about developing the sandwich ELISA using the fenobody and RANbody as pairing reagents. Results: A platform for developing a sandwich ELISA utilizing fenobody as the capture antibody and RANbody as the detection antibody was firstly designed in the study. Newcastle disease virus (NDV) was selected as the antigen, from which 13 NDV-specific nanobodies were screened from an immunized Bactrian camel. Then, 5 nanobodies were selected to produce fenobodies and RANbodies. The best pairing of fenobodies (NDV-fenobody-4, 800 ng/well) and RANbodies (NDV-RANbody-49, 1:10) was determined to develop the sandwich ELISA for detecting NDV. The detection limits of the assay were determined to be 2 2 of hemagglutination (HA) titers and 10 ng of purified NDV particles. Compared with two commercial assays, the developed assay shows higher sensitivity and specificity. Meanwhile, it exhibits 98.7% agreement with the HA test and can detect the reference NDV strains belonging to Class II but not Class I. Conclusions: In the presented study, the 13 anti-NDV nanobodies binding the NDV particles were first produced. Then, for the first time, the sandwich ELISA to detect the NDV in the different samples has been developed using the fenobody and RANbody as reagents derived from the nanobodies. Considering the rapidly increasing generation of nanobodies, the platform can reduce the cost of production for the sandwich ELISA and be universally used to develop assays for detecting other antigens.

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The role of endoglin and its soluble form in pathogenesis of preeclampsia

et al. 2021

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New highly sensitive sandwich ELISA system for soluble endoglin quantification in different biological fluids

Cited by 12 publications

References 33 publications

Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

The role of endoglin and its soluble form in pathogenesis of preeclampsia

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