Abstract
A microparticle‐enhanced nephelometric immunoassay was developed for κ‐casein quantification in human milk. Together with a previously reported β‐casein comparable immunoassay, it was applied to 862 samples milk, collected from 82 mothers, to investigate the changes in casein concentrations in human milk during the first twelve weeks of lactation. κ‐casein immunoassay is sensitive (detection limit in the reaction mixture, 0.02 mg/L) and can be performed in diluted milk, excluding any interference or sample pretreatment. It allowed the quantitation of κ‐casein over a large range of concentrations (0.14–4.56 g/L) with accuracy and precision (coefficients of variation from 3 to 10%). β‐ and κ‐casein concentrations and percentages among milk total proteins increase between colostrum (2.6 g/L, 14.3% and 1.2 g/L, 6.5%, respectively) and transitional milk (4.4 g/L, 33.2% and 1.3 g/L, 9.5%), decrease at different rates from the third to the eighth week, then remain stable at least up to the end of the third month of lactation (2.7 g/L, 25.3% and 0.9 g/L, 8.5%). The β‐casein/κ‐casein ratio is higher in colostrum (0.61) than in transitional and mature milk (0.30) and could be related to a better digestibility of colostrum casein micelles by the neonate during the first days of life. J. Clin. Lab. Anal. 13:213–218, 1999. © 1999 Wiley‐Liss, Inc.
Keywords: κ‐casein, β‐casein, human milk, lactation, immunoassay, immunonephelometry, microparticle
REFERENCES
- 1. GROVES ML, GORDON WG. The major component of human casein. A protein phosphorylated at different levels. Arch Biochem Biophys 1970;140:47–51. Medline [DOI] [PubMed] [Google Scholar]
- 2. Cuillière ML, Molé C, Montagne P, Béné MC, Faure G. Measurement of β‐casein in human milk by microparticle‐enhanced nephelometric immunoassay. Food Agric Immunol 1997;9:185–192. [Google Scholar]
- 3. Shen LH, Robberecht H, Van Dael P, Deelstra H. Estimation of bioavailability of zinc and calcium from human, cow's, goat and sheep milk by an in vitro method. Biol Trace Elem Res 1995;49:107–118. Medline [DOI] [PubMed] [Google Scholar]
- 4. Azuma N, Yamauchi K, Mitsuoka T. Bifidus growth‐promoting activity of a glycomacropeptide derived from humanκ‐casein. Agric Biol Chem 1984;48:2159–2162. [Google Scholar]
- 5. Bezkorovainy A, Grohlich D, Nichols JH. Isolation of a glycopeptide fraction with lactobacillus bifidus subspecies pennsylvanicus growth‐promoting activity from whole human milk casein. Am J Clin Nutr 1979;32:1428–1432. Medline [DOI] [PubMed] [Google Scholar]
- 6. Strömqvist M, Falk P, Bergström S, et al. Human milk κ‐casein and inhibition of Helicobacter pylori adhesion to human gastric mucosa. J Pediatr Gastr Nutr 1995;21:288–296.Medline [DOI] [PubMed] [Google Scholar]
- 7. Bergström S, Hanson L, Hernell O, Lönnerdal B, Nilsson AK, Strömqvist M. Clonning and sequencing of human κ‐casein cDNA. DNA Seq J 1992;3:245–246. [DOI] [PubMed] [Google Scholar]
- 8. Brignon G, Chtourou A, Ribadeau‐Dumas B. Preparation and amino acid sequence of human κ‐casein. FEBS Lett 1985;188:48–54. Medline [DOI] [PubMed] [Google Scholar]
- 9. Azuma N, Kaminogawa S, Yamauchi K. Reconstitution of human casein micelle and its properties. Agric Biol Chem 1985;49:2655–2660. [Google Scholar]
- 10. Dev BC, Sood SM, DeWind S, Slattery CW. Characterization of human κ‐casein purified by FPLC. Prep Biochem 1993;23:389–407. Medline [DOI] [PubMed] [Google Scholar]
- 11. Dev BC, Sood SM, DeWind S, Slattery CW. κ‐casein and β‐caseins in human milk micelles: Structural studies. Arch Biochem Biophys 1994;314:329–336. Medline [DOI] [PubMed] [Google Scholar]
- 12. Carroll RJ, Basch JJ, Phillips JG, Farrell HM. Ultrastructural and biochemical investigations of mature human milk. Food Microstruct 1985;4:321–323. [Google Scholar]
- 13. Farrell HM, Pessen H, Brown EM, Kamosinski TF. Structural insights into the bovine casein micelle: Small angle X‐ray scattering studies and correlations with spectroscopy. J Dairy Sci 1990;73:3592–3601. [Google Scholar]
- 14. Cavaletto M, Cantisani A, Napolitano L, et al. Comparative study of casein content in human colostrum and milk. Milchwissenschaft 1994;49:303–305. [Google Scholar]
- 15. Kroening TA, Pradip Mukerji MS, Hards RG. Analysis of beta‐casein and its phosphoforms in human milk. Nutr Res 1998;18:1175–1186. [Google Scholar]
- 16. Montagne P, Laroche P, Bessou TH, Cuillière ML, Varcin P, Duheille J. Measurement of eleven serum proteins by microparticle‐enhanced nephelometric immunoassay. J Clin Chem Clin Biochem 1992;30:217–222. [DOI] [PubMed] [Google Scholar]
- 17. Collard‐Bovy C, Marchal E, Humbert G, et al. Microparticle‐enhanced nephelometric immunoassay: I‐Measurement of alphas‐ and kappa‐caseins. J Dairy Sci 1991;74:3695–3701. [Google Scholar]
- 18. El Bari N, Montagne P, Humbert G, et al. Development of a microparticle‐enhanced nephelometric immunoassay for the quantification of beta‐casein in milk. Food Agric Immunol 1991;3:63–71. [Google Scholar]
- 19. Montagne P, Varcin P, Cuillière ML, Duheille J. Microparticle‐enhanced nephelometric immunoassay with microsphere‐antigen conjugates. Bioconjug Chem 1992;3:187–193. Medline [DOI] [PubMed] [Google Scholar]
- 20. Marchand J, Varcin P, Riochet D, et al. Synthesis of new hydrophilic microspheres: Optimized carriers for microparticle‐enhanced nephelometric immunoassay. Biopolymers 1992;32:971–980. Medline [DOI] [PubMed] [Google Scholar]
- 21. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein‐dye binding. Anal Biochem 1976;72:248–254. Medline [DOI] [PubMed] [Google Scholar]
- 22. Altschul SF, Warren G, Webb M, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol 1990;215:403–410. Medline [DOI] [PubMed] [Google Scholar]
- 23. Azuma N, Kaminogawa S, Yamauchi K. Properties of glycomacropeptide and para‐k‐casein derived from human k‐casein and comparison of human and bovine k‐caseins as to susceptibility to chymosin and pepsin. Agric Biol Chem 1984;48:2025–2031. [Google Scholar]
- 24. Mercier JC, Chobert, JM. Comparative study of the amino acid sequence of the caseinomacropeptides from seven species. FEBS Lett 1976;72:208–214. [DOI] [PubMed] [Google Scholar]
- 25. Otani H, Higashiyama S, Tokita F. Antigenic reactivity of human casein with antiserum to bovine β‐casein. Milchwissenschaft 1984;39:65–67. [Google Scholar]
- 26. Yvon M, Pélissier JP. Characterization and kinetics of evacuation of peptides resulting from casein hydrolysis in the stomach of the calf. J Agric Food Chem 1987;35:148–156. [Google Scholar]
