Generation of an anti-NAGase single chain antibody and its application in a biosensor-based assay for the detection of NAGase in milk.
J Immunol Methods
; 364(1-2): 14-20, 2011 Feb 01.
Article
em En
| MEDLINE
| ID: mdl-20883698
ABSTRACT
Bovine mastitis, an inflammation of the mammary gland in cows, is a major challenge for the dairy industry worldwide as it lowers milk yield, reduces milk quality and increases overall production costs. Early diagnosis is of the utmost importance. N-acetyl-ß-D-glucosaminidase (NAGase) is an enzyme released into milk during inflammation and acts as an early indicator of mastitis. This paper describes the selection of anti-NAGase single chain fragment variable antibodies (scFv) from naïve human antibody libraries and their incorporation into an automated optical biosensor-based immunoassay to detect NAGase in milk. The scFv with the highest affinity for NAGase was first characterized by inhibition ELISA, followed by further evaluation using a surface plasmon resonance platform. Purified NAGase was immobilized on the surface of a CM5 chip and spiked NAGase milk samples were analyzed. The limit of detection for the assay for the assay was determined as 1µg/ml.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Acetilglucosaminidase
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Biomarcadores
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Leite
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Anticorpos de Cadeia Única
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Mastite Bovina
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Irlanda