Imunoradioassay validation to plasmatic bovine leptin quantification
Autor(a) principal: | |
---|---|
Data de Publicação: | 2010 |
Outros Autores: | |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Brazilian Journal of Veterinary Research and Animal Science |
Texto Completo: | https://www.revistas.usp.br/bjvras/article/view/26843 |
Resumo: | Due necessity of better understanding leptin and reproduction relations, a specific radioimmunoassay (RIA) to bovine leptin was validated. First, an antibody production protocol was developed using recombinant equine leptin inoculated in a rabbit, that results in 28,05% of maximum binding (MB) 105 days after the protocol beginning. The tests of validations verified parallelism between standard curve and dilutions of high and low controls (P < 0,01). Antibody against equine leptin showed specificity to bovine leptin (P < 0,01). The recuperation tax of bovine leptin by antibody against recombinant equine leptin was from 98,4 to 101,6% (P < 0, 01). When the samples were stored in ambient temperature or refrigerated to 4 °C, ligation stability was verified (P >; 0,2), however, temperatures above 37 °C impaired the bovine leptin recuperation. The use of assay buffer with or without bovine plasma did not show any difference (P >; 0,3). These results showed that the antibody produced in rabbit against equine leptin were able to detect plasmatic bovine leptin, and that the RIA to bovine leptin quantification had adequate characteristics to the development of a valid assay. |
id |
USP-49_65d110b789ed673f8cf899a73232b664 |
---|---|
oai_identifier_str |
oai:revistas.usp.br:article/26843 |
network_acronym_str |
USP-49 |
network_name_str |
Brazilian Journal of Veterinary Research and Animal Science |
repository_id_str |
https://www.revistas.usp.br/bjvras/index |
spelling |
Imunoradioassay validation to plasmatic bovine leptin quantificationValidação de radioimunoensaio para quantificação de leptina plasmática bovinaLeptinaAnticorpoRadioimunoensaioValidaçãoLeptinAntibodyRadioimmunoassayValidationDue necessity of better understanding leptin and reproduction relations, a specific radioimmunoassay (RIA) to bovine leptin was validated. First, an antibody production protocol was developed using recombinant equine leptin inoculated in a rabbit, that results in 28,05% of maximum binding (MB) 105 days after the protocol beginning. The tests of validations verified parallelism between standard curve and dilutions of high and low controls (P < 0,01). Antibody against equine leptin showed specificity to bovine leptin (P < 0,01). The recuperation tax of bovine leptin by antibody against recombinant equine leptin was from 98,4 to 101,6% (P < 0, 01). When the samples were stored in ambient temperature or refrigerated to 4 °C, ligation stability was verified (P >; 0,2), however, temperatures above 37 °C impaired the bovine leptin recuperation. The use of assay buffer with or without bovine plasma did not show any difference (P >; 0,3). These results showed that the antibody produced in rabbit against equine leptin were able to detect plasmatic bovine leptin, and that the RIA to bovine leptin quantification had adequate characteristics to the development of a valid assay.Devido à necessidade de compreender melhor as interações entre leptina e reprodução, um RIA específico para a leptina bovina foi validado. Primeiro, um protocolo para produção de anticorpos foi desenvolvido por meio da inoculação de leptina recombinante equina em um coelho, que resultou em 28,05% de ligação máxima (MB) 105 dias após o inicio do protocolo. Os testes de validação verificaram paralelismo entre a curva-padrão e as diluições dos controles alto e baixo (p; 0,2), no entanto, temperaturas acima de 37 °C interferiram negativamente na recuperação da leptina bovina. O uso do tampão de ensaio com ou sem a adição de plasma não apresentou diferenças (p >; 0,3). Esses resultados demonstraram que o anticorpo produzido em coelho contra leptina equina foi capaz de detectar a leptina plasmática bovina, e que o RIA para a quantificação da leptina bovina apresentou características adequadas para o desenvolvimento de um ensaio válido.Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia2010-02-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/bjvras/article/view/2684310.11606/issn.1678-4456.bjvras.2010.26843Brazilian Journal of Veterinary Research and Animal Science; Vol. 47 Núm. 1 (2010); 05-12Brazilian Journal of Veterinary Research and Animal Science; Vol. 47 No. 1 (2010); 05-12Brazilian Journal of Veterinary Research and Animal Science; v. 47 n. 1 (2010); 05-12Brazilian Journal of Veterinary Research and Animal Science; V. 47 N. 1 (2010); 05-121678-44561413-9596reponame:Brazilian Journal of Veterinary Research and Animal Scienceinstname:Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)instacron:USPporhttps://www.revistas.usp.br/bjvras/article/view/26843/28626Beltran, Maria PaulaNogueira, Guilherme de Paulainfo:eu-repo/semantics/openAccess2020-06-23T04:11:06Zoai:revistas.usp.br:article/26843Revistahttps://www.revistas.usp.br/bjvrasPUBhttps://www.revistas.usp.br/bjvras/oaibjvras@usp.br1413-95961413-9596opendoar:https://www.revistas.usp.br/bjvras/index2023-01-12T16:43:07.191761Brazilian Journal of Veterinary Research and Animal Science - Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)false |
dc.title.none.fl_str_mv |
Imunoradioassay validation to plasmatic bovine leptin quantification Validação de radioimunoensaio para quantificação de leptina plasmática bovina |
title |
Imunoradioassay validation to plasmatic bovine leptin quantification |
spellingShingle |
Imunoradioassay validation to plasmatic bovine leptin quantification Beltran, Maria Paula Leptina Anticorpo Radioimunoensaio Validação Leptin Antibody Radioimmunoassay Validation |
title_short |
Imunoradioassay validation to plasmatic bovine leptin quantification |
title_full |
Imunoradioassay validation to plasmatic bovine leptin quantification |
title_fullStr |
Imunoradioassay validation to plasmatic bovine leptin quantification |
title_full_unstemmed |
Imunoradioassay validation to plasmatic bovine leptin quantification |
title_sort |
Imunoradioassay validation to plasmatic bovine leptin quantification |
author |
Beltran, Maria Paula |
author_facet |
Beltran, Maria Paula Nogueira, Guilherme de Paula |
author_role |
author |
author2 |
Nogueira, Guilherme de Paula |
author2_role |
author |
dc.contributor.author.fl_str_mv |
Beltran, Maria Paula Nogueira, Guilherme de Paula |
dc.subject.por.fl_str_mv |
Leptina Anticorpo Radioimunoensaio Validação Leptin Antibody Radioimmunoassay Validation |
topic |
Leptina Anticorpo Radioimunoensaio Validação Leptin Antibody Radioimmunoassay Validation |
description |
Due necessity of better understanding leptin and reproduction relations, a specific radioimmunoassay (RIA) to bovine leptin was validated. First, an antibody production protocol was developed using recombinant equine leptin inoculated in a rabbit, that results in 28,05% of maximum binding (MB) 105 days after the protocol beginning. The tests of validations verified parallelism between standard curve and dilutions of high and low controls (P < 0,01). Antibody against equine leptin showed specificity to bovine leptin (P < 0,01). The recuperation tax of bovine leptin by antibody against recombinant equine leptin was from 98,4 to 101,6% (P < 0, 01). When the samples were stored in ambient temperature or refrigerated to 4 °C, ligation stability was verified (P >; 0,2), however, temperatures above 37 °C impaired the bovine leptin recuperation. The use of assay buffer with or without bovine plasma did not show any difference (P >; 0,3). These results showed that the antibody produced in rabbit against equine leptin were able to detect plasmatic bovine leptin, and that the RIA to bovine leptin quantification had adequate characteristics to the development of a valid assay. |
publishDate |
2010 |
dc.date.none.fl_str_mv |
2010-02-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.revistas.usp.br/bjvras/article/view/26843 10.11606/issn.1678-4456.bjvras.2010.26843 |
url |
https://www.revistas.usp.br/bjvras/article/view/26843 |
identifier_str_mv |
10.11606/issn.1678-4456.bjvras.2010.26843 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
https://www.revistas.usp.br/bjvras/article/view/26843/28626 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia |
publisher.none.fl_str_mv |
Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia |
dc.source.none.fl_str_mv |
Brazilian Journal of Veterinary Research and Animal Science; Vol. 47 Núm. 1 (2010); 05-12 Brazilian Journal of Veterinary Research and Animal Science; Vol. 47 No. 1 (2010); 05-12 Brazilian Journal of Veterinary Research and Animal Science; v. 47 n. 1 (2010); 05-12 Brazilian Journal of Veterinary Research and Animal Science; V. 47 N. 1 (2010); 05-12 1678-4456 1413-9596 reponame:Brazilian Journal of Veterinary Research and Animal Science instname:Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP) instacron:USP |
instname_str |
Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Brazilian Journal of Veterinary Research and Animal Science |
collection |
Brazilian Journal of Veterinary Research and Animal Science |
repository.name.fl_str_mv |
Brazilian Journal of Veterinary Research and Animal Science - Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP) |
repository.mail.fl_str_mv |
bjvras@usp.br |
_version_ |
1797051559888027648 |