Development of enzyme immunoassay for detecting I and II types of shiga-like toxins

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Introduction. The aim of the work was development of enzyme immunoassay for detecting I and II types of shiga-like toxins and assessment of it diagnostic properties. 

Materials and methods. For the research, we used hybridomas producing monoclonal antibodies to shiga-like toxins of types I and II, obtained at the branch of the Federal State Budgetary Institution “48 Central Research Institute” of the Ministry of Defense of Russian Federation (Kirov); BALB/c mice; shiga-like toxins of types I and II. Hybridoma cells were cultured in culture flasks and in the peritoneal cavity of BALB/c mice. Monoclonal antibodies were isolated from ascitic fluids by precipitation with a saturated solution of ammonium sulfate, followed by purification by ion exchange chromatography. The obtained preparations of monoclonal antibodies were used to develop enzyme immunoassay for the detection of shiga-like toxins of types I and II. Specific components of enzyme immunoassay were freeze-dried in a protective environment.

Results. As a result of research, preparative quantities of monoclonal antibodies against I and II types of shiga-like toxins were obtained and purified; selection of monoclonal antibodies for sorption on the solid phase and for the synthesis of immunoperoxidase conjugates was carried out. 

Conclusion. Experimental enzyme immunoassay allowing to identify 1 ng/ml I and II types of shiga-like toxins in «sandwich»-ELISA was developed.

Sobre autores

Galina Kuklina

Affiliated Branch of the Federal State Budgetary Institution «48th Central Research Institute» of the Ministry of Defense of the Russian Federation

Autor responsável pela correspondência
Email: noemail@neicon.ru
ORCID ID: 0000-0002-5429-6295

Кандидат биологических наук, младший научный сотрудник научно-исследовательского
отдела Филиала ФГБУ 48 ЦНИИ Минобороны РФ, 610000, г. Киров, Россия

Rússia

Denis Pechenkin

Affiliated Branch of the Federal State Budgetary Institution «48th Central Research Institute» of the Ministry of Defense of the Russian Federation

Email: noemail@neicon.ru
ORCID ID: 0000-0001-8277-3573

Ph.D. in Medicine, Head of Research Department Affiliated Branch of the Federal State Budgetary Institution «48th Central Research Institute» of the Ministry of Defense of the Russian Federation, Kirov, 610000, Russian Federation, 610000, Kirov, Russia.

e-mail: 23527@mil.ru

Rússia

Sergei Ipatov

Affiliated Branch of the Federal State Budgetary Institution «48th Central Research Institute» of the Ministry of Defense of the Russian Federation

Email: noemail@neicon.ru
ORCID ID: 0000-0002-2881-4730

Младший научный сотрудник научно-исследовательского отдела Филиала ФГБУ 48 ЦНИИ Минобороны РФ, 610000, г. Киров, Россия.

Rússia

Andrei Eremkin

Affiliated Branch of the Federal State Budgetary Institution «48th Central Research Institute» of the Ministry of Defense of the Russian Federation

Email: noemail@neicon.ru
ORCID ID: 0000-0003-3133-1921

Кандидат биологических наук, начальник группы – заместитель начальника научно-исследовательского отдела Филиала ФГБУ 48 ЦНИИ Минобороны РФ, 610000, г. Киров, Россия.

Rússia

Aleksei Kytmanov

Affiliated Branch of the Federal State Budgetary Institution «48th Central Research Institute» of the Ministry of Defense of the Russian Federation

Email: noemail@neicon.ru
ORCID ID: 0000-0003-0277-2226

Кандидат биологических наук, научный сотрудник научно-исследовательского отдела Филиала ФГБУ 48 ЦНИИ Минобороны РФ, 610000, г. Киров, Россия.

Rússia

Oksana Tikhvinskaya

Affiliated Branch of the Federal State Budgetary Institution «48th Central Research Institute» of the Ministry of Defense of the Russian Federation

Email: noemail@neicon.ru
ORCID ID: 0000-0001-8368-9648

Кандидат биологических наук, младший научный сотрудник научно-исследовательского отдела Филиала ФГБУ 48 ЦНИИ Минобороны РФ, 610000, г. Киров, Россия.

Rússia

Bibliografia

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Declaração de direitos autorais © Kuklina G.V., Pechenkin D.V., Ipatov S.S., Eremkin A.V., Kytmanov A.A., Tikhvinskaya O.V., 2021


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