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https://www.arca.fiocruz.br/handle/icict/46087
MAGNETIC BEAD-BASED IMMUNOASSAY ALLOWS RAPID, INEXPENSIVE, AND QUANTITATIVE DETECTION OF HUMAN SARS-COV‑2 ANTIBODIES
Enzyme-Linked Immunosorbent Assay
Immunological Models
Antibodies
Methods
Ensayo de Inmunoadsorción Enzimática
Modelos Inmunológicos
Anticuerpos
Métodos
Ensaio de Imunoadsorção Enzimática
Modelos Imunológicos
Anticorpos
Métodos
Author
Huergo, Luciano F.
Selim, Khaled A.
Conzentino, Marcelo S.
Gerhardt, Edileusa C. M.
Santos, Adrian R. S.
Wagner, Berenike
Alford, Janette T.
Deobald, Nelli
Pedrosa, Fabio O.
Souza, Emanuel Maltempi de
Nogueira, Meri B.
Raboni, Sônia M.
Souto, Dênio
Rego, Fabiane G. M.
Zanette, Dalila Lucíola
Aoki, Mateus Nóbrega
Nardin, Jeanine M.
Fornazari, Bruna
Morales, Hugo M. P.
Borges, Vânia A.
Nelde, Annika
Walz, Juliane S.
Becker, Matthias
Schneiderhan-Marra, Nicole
Rothbauer, Ulrich
Reis, Rodrigo A.
Forchhammer, Karl
Selim, Khaled A.
Conzentino, Marcelo S.
Gerhardt, Edileusa C. M.
Santos, Adrian R. S.
Wagner, Berenike
Alford, Janette T.
Deobald, Nelli
Pedrosa, Fabio O.
Souza, Emanuel Maltempi de
Nogueira, Meri B.
Raboni, Sônia M.
Souto, Dênio
Rego, Fabiane G. M.
Zanette, Dalila Lucíola
Aoki, Mateus Nóbrega
Nardin, Jeanine M.
Fornazari, Bruna
Morales, Hugo M. P.
Borges, Vânia A.
Nelde, Annika
Walz, Juliane S.
Becker, Matthias
Schneiderhan-Marra, Nicole
Rothbauer, Ulrich
Reis, Rodrigo A.
Forchhammer, Karl
Affilliation
Universidade Federal do Paraná. Setor Litoral. Matinhos. PR, Brasil.
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany.
Universidade Federal do Paraná. Setor Litoral. Matinhos. PR, Brasil.
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany .
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany.
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany.
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Complexo Hospital de Clínicas. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Complexo Hospital de Clínicas. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Departamento de Química. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Programa de Pós-Graduação em Ciências Farmacêuticas. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Hospital Erasto Gaertner. Curitiba, PR, Brasil.
Hospital Erasto Gaertner. Curitiba, PR, Brasil.
Hospital Erasto Gaertner. Curitiba, PR, Brasil.
Secretaria Municipal de Saúde de Guaratuba. Guaratuba, PR. Brasil.
Clinical Collaboration Unit Translational Immunology. German Cancer Consortium (DKTK). / Department of Internal Medicine. University Hospital Tübingen.Tübingen, Germany. / Department of Immunology. Institute for Cell Biology and Cluster of Excellence. Tübingen University. Tübingen, Germany.
Clinical Collaboration Unit Translational Immunology. German Cancer Consortium (DKTK). / Department of Internal Medicine and Departament of Hematooogy, Oncology, Clinical Immunology and Rheumatology. University Hospital Tübingen.Tübingen, Germany. / Department of Immunology. Institute for Cell Biology and Cluster of Excellence. Tübingen University. Tübingen, Germany.
Tübingen University. Institute of Natural and Medical Sciences. Tübingen, Germany.
Tübingen University. Institute of Natural and Medical Sciences. Tübingen, Germany.
Tübingen University. Institute of Natural and Medical Sciences. Tübingen, Germany. / Tübingen University. Pharmaceutical Biotechnology. Tübingen, Germany.
Universidade Federal do Paraná. Setor Litoral. Matinhos. PR, Brasil.
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany.
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany.
Universidade Federal do Paraná. Setor Litoral. Matinhos. PR, Brasil.
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany .
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany.
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany.
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Departamento de Bioquímica e Biologia Molecular. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Complexo Hospital de Clínicas. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Complexo Hospital de Clínicas. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Departamento de Química. Curitiba, PR, Brasil.
Universidade Federal do Paraná. Programa de Pós-Graduação em Ciências Farmacêuticas. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Hospital Erasto Gaertner. Curitiba, PR, Brasil.
Hospital Erasto Gaertner. Curitiba, PR, Brasil.
Hospital Erasto Gaertner. Curitiba, PR, Brasil.
Secretaria Municipal de Saúde de Guaratuba. Guaratuba, PR. Brasil.
Clinical Collaboration Unit Translational Immunology. German Cancer Consortium (DKTK). / Department of Internal Medicine. University Hospital Tübingen.Tübingen, Germany. / Department of Immunology. Institute for Cell Biology and Cluster of Excellence. Tübingen University. Tübingen, Germany.
Clinical Collaboration Unit Translational Immunology. German Cancer Consortium (DKTK). / Department of Internal Medicine and Departament of Hematooogy, Oncology, Clinical Immunology and Rheumatology. University Hospital Tübingen.Tübingen, Germany. / Department of Immunology. Institute for Cell Biology and Cluster of Excellence. Tübingen University. Tübingen, Germany.
Tübingen University. Institute of Natural and Medical Sciences. Tübingen, Germany.
Tübingen University. Institute of Natural and Medical Sciences. Tübingen, Germany.
Tübingen University. Institute of Natural and Medical Sciences. Tübingen, Germany. / Tübingen University. Pharmaceutical Biotechnology. Tübingen, Germany.
Universidade Federal do Paraná. Setor Litoral. Matinhos. PR, Brasil.
Interfaculty Institute for Microbiology and Infection Medicine. Eberhard Karls University of Tübingen. Tübingen, Germany.
Abstract
Immunological methods to detect SARS-CoV-2 seroconversion in humans are important to track COVID-19 cases and the humoral response to SARS-CoV-2 infections and immunization to future vaccines. The aim of this work was to develop a simple chromogenic magnetic bead-based immunoassay which allows rapid, inexpensive, and quantitative detection of human antibodies against SARS-CoV-2 in serum, plasma, or blood. Recombinant 6xHis-tagged SARS-CoV-2 Nucleocapsid protein was mobilized on the surface of Ni2+ magnetic beads and challenged with serum or blood samples obtained from controls or COVID-19 cases. The beads were washed, incubated with anti-human IgG-HPR
conjugate, and immersed into a solution containing a chromogenic HPR substrate. Bead transfer and homogenization between solutions was aided by a simple low-cost device. The method was validated by two independent laboratories, and the performance to detect SARS-CoV-2 seroconversion in humans was in the same range as obtained using the gold standard immunoassays ELISA and Luminex, though requiring only a fraction of consumables, instrumentation, time to deliver results, and volume of sample. Furthermore, the results obtained with the method described can be visually interpreted without compromising accuracy as demonstrated by validation at a point-of-care unit. The magnetic bead immunoassay throughput can be customized on demand and is readily adapted to be used with any other 6xHis tagged protein or peptide as antigen to track other diseases.
Keywords
Coronavirus InfectionsEnzyme-Linked Immunosorbent Assay
Immunological Models
Antibodies
Methods
Keywords in Spanish
Infecciones por CoronavirusEnsayo de Inmunoadsorción Enzimática
Modelos Inmunológicos
Anticuerpos
Métodos
DeCS
Infecções por CoronavirusEnsaio de Imunoadsorção Enzimática
Modelos Imunológicos
Anticorpos
Métodos
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