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Microparticle-enhanced nephelometric immunoassay for caseinomacropeptide in milk

Published online by Cambridge University Press:  01 June 2009

Christine Prin ,
Nezha El Bari ,
Paul Montagne ,
Marie-Louise Cuilliere ,
Marie-Christine Bene ,
Gilbert Faure ,
Gerard Humbert  and
Guy Linden
Christine Prin
Affiliation:
Laboratoire d'Immunologie, Faculté de Médecine, UFR Sciences Médicales, BP 184, F-54505 Vandœuvre-les-Nancy CedexFrance
Nezha El Bari
Affiliation:
Laboratoire d'Immunologie, Faculté de Médecine, UFR Sciences Médicales, BP 184, F-54505 Vandœuvre-les-Nancy CedexFrance
Paul Montagne
Affiliation:
Laboratoire d'Immunologie, Faculté de Médecine, UFR Sciences Médicales, BP 184, F-54505 Vandœuvre-les-Nancy CedexFrance
Marie-Louise Cuilliere
Affiliation:
Laboratoire d'Immunologie, Faculté de Médecine, UFR Sciences Médicales, BP 184, F-54505 Vandœuvre-les-Nancy CedexFrance
Marie-Christine Bene
Affiliation:
Laboratoire d'Immunologie, Faculté de Médecine, UFR Sciences Médicales, BP 184, F-54505 Vandœuvre-les-Nancy CedexFrance
Gilbert Faure
Affiliation:
Laboratoire d'Immunologie, Faculté de Médecine, UFR Sciences Médicales, BP 184, F-54505 Vandœuvre-les-Nancy CedexFrance
Gerard Humbert
Affiliation:
Laboratoire de Biochimie Appliquée associé à INRA, Faculté des Sciences, Université Henri Poincaré-Nancy 1, BP 239, F-54506 Vandœuvre-les-Nancy Cedex, France
Guy Linden
Affiliation:
Laboratoire de Biochimie Appliquée associé à INRA, Faculté des Sciences, Université Henri Poincaré-Nancy 1, BP 239, F-54506 Vandœuvre-les-Nancy Cedex, France
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Summary

A microparticle-enhanced nephelometric immunoassay has been developed for the determination of caseinomacropeptide (CMP) in bovine milk. It is based on the nephelometric quantification of the competitive immunoagglutination of a microparticle–CMP conjugate with an anti-κ-casein (κ-CN) antiserum. This one step immunoassay was sensitive (detection limit in reaction mixture, 16μg/l), accurate (linear recovery of CMP in dilution overloading) and reproducible (CV 7–14% for within and between run precision). Because of the specificity of the polyclonal antiserum used, it was necessary to separate CMP from κ-CN by ultrafiltration before the quantification of bovine milk CMP. Under the conditions of milk ultrafiltration used, κ-CN was entirely retained (> 99·5%) but the concentration of CMP measured in milk ultrafiltrates was underestimated (by ∼25%) compared with its concentration in whole milk. Microparticle-enhanced nephelometric immunoassay of CMP, with a calibration range from 0·32 to 20 mg/1 for 20- fold diluted milk ultrafiltrate, allowed contamination of bovine milk by rennet whey as low as 5 ml/1 to be detected. Applied to ultrafiltrates from milk stored at 4 °C, this immunoassay also detected proteolysis of κ-CN not revealed by measurement of κ-CN concentration in milk. A statistical lower limit of 3·21 mg/1 was determined as the increase in CMP concentration in milk ultrafiltrates that indicated probable κ-CN proteolysis in the milk sample. Previously demonstrated to be an easy to perform method for assaying the main proteins of bovine milk, microparticle-enhanced nephelometric immunoassay thus also appeared to be appropriate to quantify CMP so as to detect slight contamination of milk by whey and to indicate the proteolysis of κ-CN during milk storage at low temperature.

Type
Original Articles
Information
Journal of Dairy Research , Volume 63 , Issue 1 , February 1996 , pp. 73 - 81
Copyright
Copyright © Proprietors of Journal of Dairy Research 1996

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