Peer-Reviewed Journal Details
Mandatory Fields
Welbeck K.;Leonard P.;Gilmartin N.;Byrne B.;Viguier C.;Arora S.;O'Kennedy R.
2011
February
Journal of Immunological Methods
Generation of an anti-NAGase single chain antibody and its application in a biosensor-based assay for the detection of NAGase in milk
Published
19 ()
Optional Fields
Immunoassay NAGase ScFv SPR
364
1-2
14
20
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-β-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 1 μg/ml. © 2010 Elsevier B.V.
0022-1759
10.1016/j.jim.2010.09.019
Grant Details