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Multiplexing of Human Preterm and Term Milk Cytokines
Author(s) -
Groer Maureen Edith,
Williams S. Nicole,
Kane Bradley Patrick
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.629.7
Subject(s) - colostrum , multiplexing , medicine , matrix (chemical analysis) , immune system , immunology , cytokine , computer science , chemistry , antibody , chromatography , telecommunications
The first objective was to use an 18‐plex cytokine panel to compare preterm and term colostrum collected at day 1 postpartum. After observing matrix effects, we worked to develop a milk assay matrix for Luminex and MagPix technologies. After matrix development proved satisfactory, we have progressed to a longitudinal study of immunobiology of milk from 100 mothers of Very Low Birth Weight (VLBW) infants. We multiplexed milk whey using bead based methods and performed linear spiking experiments. We used a variety of matrix solutions and these matrices did not give reliable results. When we used serum matrix for controls and standards, and added serum matrix to milk samples, the spiking experiments were successful and showed linearity. Based on our preliminary experiments on preterm and term colostrum and the matrix solution, we designed a plex of 8 cytokines that differed in preterm compared to term colostrum (IL‐4, IL‐6, IL‐8, IL‐10, IP‐10, MIP‐1α, IP‐10, MCP‐1, and TNFα). We are now measuring milk cytokines from 100 mothers of VLBW infants across 6 weeks of the NICU stay. From these data we will describe the trajectory of human milk immunobiology over the first six weeks postpartum. Multiplexing provides excellent screening for important immune molecules. However there are no plexes specifically for milk. We have developed an assay for multiplexing using serum matrix that is sensitive and reliable. Funded by 5R21NR013094–02