Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis

Segovia, Paula - Quinteros, Pedro - Morais, Sergi - Echaides, Cesar - Maquieira, Ángel - Lassabe, Gabriel - Gonzalez Sapienza, Gualberto

Resumen:

Commonly, serological immunoassays and diagnostic kits include reference standard reagents (calibrators) that contain the specific antibodies to be measured, which are used for the quantification of the unknown antibody contained in the sample. However, in some cases, such as the diagnosis of allergies or autoimmune diseases, it is often difficult to have sufficient quantities of these reference standards, and there are also limitations to their lot-to-lot reproducibility and standardization over time. To overcome this difficulty, this work introduces the use of surrogate recombinant calibrators formulated on the basis of two single domain antibodies (nanobodies) combined through a short peptide linker to produce a recombinant bispecific construct. One of the nanobodies binds to the cognate antigen of the target antibody and the second is specific for the paratope of the secondary detecting antibody. The bispecific nanobody inherits the outstanding properties of stability and low cost production by bacterial fermentation of the parent nanobodies, and once calibrated against the biological reference standard, it can be reproduced indefinitely from its sequence in a highly standardized manner. As proof of concept, we present the generation and characterization of two bispecific calibrators with potential application to the diagnosis of allergy against the antibiotics aztreonam and amoxicillin.

Detalles Bibliográficos
2022
INMUNOLOGIA
PEPTIDOS
PROTEINAS
BIOPOLIMEROS
AGENTES ANTIMICROBIANOS
Inglés
Universidad de la República
COLIBRI
https://hdl.handle.net/20.500.12008/37025
Acceso abierto
Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)
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author Segovia, Paula
author2 Quinteros, Pedro
Morais, Sergi
Echaides, Cesar
Maquieira, Ángel
Lassabe, Gabriel
Gonzalez Sapienza, Gualberto
author2_role author
author
author
author
author
author
author_facet Segovia, Paula
Quinteros, Pedro
Morais, Sergi
Echaides, Cesar
Maquieira, Ángel
Lassabe, Gabriel
Gonzalez Sapienza, Gualberto
author_role author
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dc.contributor.filiacion.none.fl_str_mv Segovia Paula. Universidad de la República (Uruguay). Facultad de Química. DEPBIO. Cátedra de Inmunología. Instituto de Higiene
Quinteros Pedro. Universidad Politécnica de Valencia (España). Departamento de Química
Morais Sergi. Universidad Politécnica de Valencia (España). Departamento de Química
Echaides Cesar. Intendencia de Montevideo (Uruguay). Parque Lecoq
Maquieira Ángel. Universidad Politécnica de Valencia (España). Departamento de Química
Lassabe Gabriel. Universidad de la República (Uruguay) Facultad de Química. DEPBIO. Cátedra de Inmunología. Instituto de Higiene
Gonzalez Sapienza Gualberto. Universidad de la República (Uruguay) Facultad de Química. DEPBIO. Cátedra de Inmunología. Instituto de Higiene
dc.creator.none.fl_str_mv Segovia, Paula
Quinteros, Pedro
Morais, Sergi
Echaides, Cesar
Maquieira, Ángel
Lassabe, Gabriel
Gonzalez Sapienza, Gualberto
dc.date.accessioned.none.fl_str_mv 2023-05-09T15:44:20Z
dc.date.available.none.fl_str_mv 2023-05-09T15:44:20Z
dc.date.issued.none.fl_str_mv 2022
dc.description.abstract.none.fl_txt_mv Commonly, serological immunoassays and diagnostic kits include reference standard reagents (calibrators) that contain the specific antibodies to be measured, which are used for the quantification of the unknown antibody contained in the sample. However, in some cases, such as the diagnosis of allergies or autoimmune diseases, it is often difficult to have sufficient quantities of these reference standards, and there are also limitations to their lot-to-lot reproducibility and standardization over time. To overcome this difficulty, this work introduces the use of surrogate recombinant calibrators formulated on the basis of two single domain antibodies (nanobodies) combined through a short peptide linker to produce a recombinant bispecific construct. One of the nanobodies binds to the cognate antigen of the target antibody and the second is specific for the paratope of the secondary detecting antibody. The bispecific nanobody inherits the outstanding properties of stability and low cost production by bacterial fermentation of the parent nanobodies, and once calibrated against the biological reference standard, it can be reproduced indefinitely from its sequence in a highly standardized manner. As proof of concept, we present the generation and characterization of two bispecific calibrators with potential application to the diagnosis of allergy against the antibiotics aztreonam and amoxicillin.
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dc.identifier.citation.es.fl_str_mv Segovia, P., Quinteros, P., Morais, S. y otros. Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis [Preprint]. Publicado en: Analytical Chemical v.94. [en línea] n° 2, 2022. pp. 1342–1349 . https://doi.org/10.1021/acs.analchem.1c04603
dc.identifier.uri.none.fl_str_mv https://hdl.handle.net/20.500.12008/37025
dc.language.iso.none.fl_str_mv en
eng
dc.rights.license.none.fl_str_mv Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
dc.source.none.fl_str_mv reponame:COLIBRI
instname:Universidad de la República
instacron:Universidad de la República
dc.subject.es.fl_str_mv INMUNOLOGIA
PEPTIDOS
PROTEINAS
BIOPOLIMEROS
AGENTES ANTIMICROBIANOS
dc.title.none.fl_str_mv Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
dc.type.es.fl_str_mv Preprint
dc.type.none.fl_str_mv info:eu-repo/semantics/preprint
dc.type.version.none.fl_str_mv info:eu-repo/semantics/submittedVersion
description Commonly, serological immunoassays and diagnostic kits include reference standard reagents (calibrators) that contain the specific antibodies to be measured, which are used for the quantification of the unknown antibody contained in the sample. However, in some cases, such as the diagnosis of allergies or autoimmune diseases, it is often difficult to have sufficient quantities of these reference standards, and there are also limitations to their lot-to-lot reproducibility and standardization over time. To overcome this difficulty, this work introduces the use of surrogate recombinant calibrators formulated on the basis of two single domain antibodies (nanobodies) combined through a short peptide linker to produce a recombinant bispecific construct. One of the nanobodies binds to the cognate antigen of the target antibody and the second is specific for the paratope of the secondary detecting antibody. The bispecific nanobody inherits the outstanding properties of stability and low cost production by bacterial fermentation of the parent nanobodies, and once calibrated against the biological reference standard, it can be reproduced indefinitely from its sequence in a highly standardized manner. As proof of concept, we present the generation and characterization of two bispecific calibrators with potential application to the diagnosis of allergy against the antibiotics aztreonam and amoxicillin.
eu_rights_str_mv openAccess
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identifier_str_mv Segovia, P., Quinteros, P., Morais, S. y otros. Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis [Preprint]. Publicado en: Analytical Chemical v.94. [en línea] n° 2, 2022. pp. 1342–1349 . https://doi.org/10.1021/acs.analchem.1c04603
instacron_str Universidad de la República
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publishDate 2022
reponame_str COLIBRI
repository.mail.fl_str_mv karina.camps@seciu.edu.uy
repository.name.fl_str_mv COLIBRI - Universidad de la República
repository_id_str 4771
rights_invalid_str_mv Licencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)
spelling Segovia Paula. Universidad de la República (Uruguay). Facultad de Química. DEPBIO. Cátedra de Inmunología. Instituto de HigieneQuinteros Pedro. Universidad Politécnica de Valencia (España). Departamento de QuímicaMorais Sergi. Universidad Politécnica de Valencia (España). Departamento de QuímicaEchaides Cesar. Intendencia de Montevideo (Uruguay). Parque LecoqMaquieira Ángel. Universidad Politécnica de Valencia (España). Departamento de QuímicaLassabe Gabriel. Universidad de la República (Uruguay) Facultad de Química. DEPBIO. Cátedra de Inmunología. Instituto de HigieneGonzalez Sapienza Gualberto. Universidad de la República (Uruguay) Facultad de Química. DEPBIO. Cátedra de Inmunología. Instituto de Higiene2023-05-09T15:44:20Z2023-05-09T15:44:20Z2022Segovia, P., Quinteros, P., Morais, S. y otros. Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis [Preprint]. Publicado en: Analytical Chemical v.94. [en línea] n° 2, 2022. pp. 1342–1349 . https://doi.org/10.1021/acs.analchem.1c04603https://hdl.handle.net/20.500.12008/37025Commonly, serological immunoassays and diagnostic kits include reference standard reagents (calibrators) that contain the specific antibodies to be measured, which are used for the quantification of the unknown antibody contained in the sample. However, in some cases, such as the diagnosis of allergies or autoimmune diseases, it is often difficult to have sufficient quantities of these reference standards, and there are also limitations to their lot-to-lot reproducibility and standardization over time. To overcome this difficulty, this work introduces the use of surrogate recombinant calibrators formulated on the basis of two single domain antibodies (nanobodies) combined through a short peptide linker to produce a recombinant bispecific construct. One of the nanobodies binds to the cognate antigen of the target antibody and the second is specific for the paratope of the secondary detecting antibody. The bispecific nanobody inherits the outstanding properties of stability and low cost production by bacterial fermentation of the parent nanobodies, and once calibrated against the biological reference standard, it can be reproduced indefinitely from its sequence in a highly standardized manner. As proof of concept, we present the generation and characterization of two bispecific calibrators with potential application to the diagnosis of allergy against the antibiotics aztreonam and amoxicillin.Submitted by Cabrera Jeniffer (jenikana@gmail.com) on 2023-05-05T12:43:26Z No. of bitstreams: 2 license_rdf: 23149 bytes, checksum: 1996b8461bc290aef6a27d78c67b6b52 (MD5) Preprint_Segovia.pdf: 933675 bytes, checksum: a1954c6753be0510b4a2a016ee669b56 (MD5)Made available in DSpace by Luna Fabiana (fabiana.luna@seciu.edu.uy) on 2023-05-09T15:44:20Z (GMT). No. of bitstreams: 2 license_rdf: 23149 bytes, checksum: 1996b8461bc290aef6a27d78c67b6b52 (MD5) Preprint_Segovia.pdf: 933675 bytes, checksum: a1954c6753be0510b4a2a016ee669b56 (MD5) Previous issue date: 2022application/pdfenengLas obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. 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- Universidad de la Repúblicafalse
spellingShingle Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
Segovia, Paula
INMUNOLOGIA
PEPTIDOS
PROTEINAS
BIOPOLIMEROS
AGENTES ANTIMICROBIANOS
status_str submittedVersion
title Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
title_full Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
title_fullStr Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
title_full_unstemmed Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
title_short Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
title_sort Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
topic INMUNOLOGIA
PEPTIDOS
PROTEINAS
BIOPOLIMEROS
AGENTES ANTIMICROBIANOS
url https://hdl.handle.net/20.500.12008/37025