Bispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosis
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.
| 2022 | |
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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) |
| _version_ | 1877554840037490688 |
|---|---|
| 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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| collection | COLIBRI |
| 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. |
| dc.format.mimetype.es.fl_str_mv | application/pdf |
| 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 |
| format | preprint |
| id | COLIBRI_e7d705e39b40bc2fd3c954bb20beb796 |
| 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 |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | COLIBRI |
| network_name_str | COLIBRI |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/37025 |
| 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. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución - No Comercial - Sin Derivadas (CC - By-NC-ND 4.0)INMUNOLOGIAPEPTIDOSPROTEINASBIOPOLIMEROSAGENTES ANTIMICROBIANOSBispecific single domain antibodies as highly standardized synthetic calibrators for immunodiagnosisPreprintinfo:eu-repo/semantics/preprintinfo:eu-repo/semantics/submittedVersionreponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaSegovia, PaulaQuinteros, PedroMorais, SergiEchaides, CesarMaquieira, ÁngelLassabe, GabrielGonzalez Sapienza, GualbertoLICENSElicense.txtlicense.txttext/plain; charset=utf-84267http://localhost:8080/xmlui/bitstream/20.500.12008/37025/5/license.txt6429389a7df7277b72b7924fdc7d47a9MD55CC-LICENSElicense_urllicense_urltext/plain; charset=utf-850http://localhost:8080/xmlui/bitstream/20.500.12008/37025/2/license_urla006180e3f5b2ad0b88185d14284c0e0MD52license_textlicense_texttext/html; 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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 |