Measurement of nine human milk proteins by nephelometric immunoassays: application to the determination of mature milk protein profile

Objectives: Microparticle-enhanced nephelometric immunoassays for six human milk proteins (β-casein, κ-casein, α-lactalbumin, serum albumin, lactoferrin, and lysozyme) and conventional immunonephelometry assays for immunoglobulin A, C3, and C4 complement proteins were developed and characterized. Design and methods: Microparticle-enhanced nephelometric immunoassays are competitive assays based on the nephelometric quantification of the inhibition of microparticle-protein conjugates immunoagglutination by the proteins to be assayed. Results: High precision (CVs ranged from 1% to 14% in within- and between-assays) and recovery (linear recovery in dilution-overloading assay) ensure a reliable determination of the main human milk proteins by single-step homogeneous nephelometric immunoassays, accurate over wide ranges of concentration. These immunoassays were easily applied to a large number of mature human milk samples (between 373 and 503 according to the proteins tested). Conclusions: The immunoassays developed could be applied to the fast determination of human milk protein profile usable for nursery milk bank and fortification.