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Biosensor assay for determination of haptoglobin in bovine milk

Åkerstedt, Maria and Björk, Lennart and Persson Waller, Karin and Sternesjö, Åse (2006). Biosensor assay for determination of haptoglobin in bovine milk. Journal of dairy research. 73, 299-305
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Abstract

Despite more than 30 years of research into mastitis diagnostics, there are few alternatives to the somatic cell count (SCC) in practical use for identification of cows with subclinical mastitis. Mastitis is not only an animal welfare problem, but also affects the yield, composition and technological properties of milk. Hence, dairy cooperatives give farmers a premium quality payment to encourage low SCC although there is no clear scientific data defining the level of SCC in bulk tank milk that is associated with additional benefits in terms of milk quality. Recent research on alternative markers for inflammatory reactions in the lactating cow, e.g. in mastitis, includes investigations of the acute phase protein, haptoglobin (Hp). So far, the content of Hp in milk has mainly been studied in relation to mastitis diagnostics, with little attention given to its importance for milk composition and technological properties. At present, Hp in milk is measured using ELISA, but this technique is not suitable for routine large-scale analysis. In recent years, optical biosensor technology has been used for automated and rapid quantitative analysis of different components in milk, but so far not for analysis of acute phase proteins. The aim of the present study was to develop a rapid and sensitive biosensor method to determine Hp in milk. An affinity sensor assay based on the interaction between Hp and haemoglobin was developed using surface plasmon resonance (SPR) biosensor technology. The assay was used to analyse Hp in composite milk samples from cows without any clinical signs of mastitis and quarter milk samples with a weak to strong reaction in the California Mastitis Test (CMT). A commercial ELISA for determination of Hp in milk was used for comparison. The limit of detection (LOD) of the biosensor assay was determined as 1.1 mg/l. Within-assay and betweenday variations were determined both with bulk tank milk spiked with human Hp and with composite milk samples containing bovine Hp. Coefficients of variation varied between 3.6 and 8.6% at concentrations between 4.0 and 12 mg/l, respectively. Agreement between the results obtained by the biosensor assay and the ELISA was satisfactory ; however, the results obtained by the biosensor were generally lower than the results obtained by the ELISA. Possible explanations for this observation are discussed.

Authors/Creators:Åkerstedt, Maria and Björk, Lennart and Persson Waller, Karin and Sternesjö, Åse
Title:Biosensor assay for determination of haptoglobin in bovine milk
Series/Journal:Journal of dairy research (1469-7629)
Year of publishing :2006
Volume:73
Page range:299-305
Publisher:Cambridge University Press
ISSN:1469-7629
Language:English
Additional Information:The original publication is available at www.cambridge.org
Publication Type:Journal article
Refereed:Yes
Full Text Status:Public
Subjects:Obsolete subject words > VETERINARY MEDICINE > Pathology
Keywords:Mastitis diagnostics, dairy cow
ID Code:3243
Department:(NL, NJ) > Dept. of Food Science (until 161231)
(VH) > Dept. of Clinical Sciences
Deposited By: Staff Epsilon
Deposited On:15 May 2008 00:00
Metadata Last Modified:02 Dec 2014 10:20

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