Abstract
Hyaluronan (HA) in human milk mediates host responses to microbial infection via TLR4- and CD44-dependent signaling. Signaling by HA is generally size specific. Because pure HA with average molecular mass (M) of 35 kDa can elicit a protective response in intestinal epithelial cells, it has been proposed that human milk HA may have a bioactive low-M component. Here we report the size distribution of HA in human milk samples from 20 unique donors. A new method for HA analysis, employing ion exchange (IEX) chromatography to fractionate HA by size and specific quantification of each size fraction by competitive enzyme-linked sorbent assay (ELSA), was developed. When separated into four fractions, milk HA with M ≤ 20 kDa, M ∼ 20 to 60 kDa, and M ∼ 60 to 110 kDa comprised averages of 1.5, 1.4, and 2.0% of the total HA, respectively. The remaining 95% was HA with M ≥ 110 kDa. Electrophoretic analysis of the higher M HA from 13 samples showed nearly identical M distributions, with an average M of approximately 440 kDa. This higher M HA component in human milk is proposed to bind to CD44 and to enhance human beta defensin 2 (HBD2) induction by the low-M HA components.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 78-88 |
| Number of pages | 11 |
| Journal | Analytical Biochemistry |
| Volume | 474 |
| DOIs | |
| State | Published - Apr 1 2015 |
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Keywords
- Electrophoresis
- Hyaluronan
- Ion exchange
- Milk
- Molecular mass
- Quantification
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
- Cell Biology
Cite this
Determination of hyaluronan molecular mass distribution in human breast milk. / Yuan, Han; Amin, Ripal; Ye, Xin; De La Motte, Carol A.; Cowman, Mary.
In: Analytical Biochemistry, Vol. 474, 01.04.2015, p. 78-88.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Determination of hyaluronan molecular mass distribution in human breast milk
AU - Yuan, Han
AU - Amin, Ripal
AU - Ye, Xin
AU - De La Motte, Carol A.
AU - Cowman, Mary
PY - 2015/4/1
Y1 - 2015/4/1
N2 - Hyaluronan (HA) in human milk mediates host responses to microbial infection via TLR4- and CD44-dependent signaling. Signaling by HA is generally size specific. Because pure HA with average molecular mass (M) of 35 kDa can elicit a protective response in intestinal epithelial cells, it has been proposed that human milk HA may have a bioactive low-M component. Here we report the size distribution of HA in human milk samples from 20 unique donors. A new method for HA analysis, employing ion exchange (IEX) chromatography to fractionate HA by size and specific quantification of each size fraction by competitive enzyme-linked sorbent assay (ELSA), was developed. When separated into four fractions, milk HA with M ≤ 20 kDa, M ∼ 20 to 60 kDa, and M ∼ 60 to 110 kDa comprised averages of 1.5, 1.4, and 2.0% of the total HA, respectively. The remaining 95% was HA with M ≥ 110 kDa. Electrophoretic analysis of the higher M HA from 13 samples showed nearly identical M distributions, with an average M of approximately 440 kDa. This higher M HA component in human milk is proposed to bind to CD44 and to enhance human beta defensin 2 (HBD2) induction by the low-M HA components.
AB - Hyaluronan (HA) in human milk mediates host responses to microbial infection via TLR4- and CD44-dependent signaling. Signaling by HA is generally size specific. Because pure HA with average molecular mass (M) of 35 kDa can elicit a protective response in intestinal epithelial cells, it has been proposed that human milk HA may have a bioactive low-M component. Here we report the size distribution of HA in human milk samples from 20 unique donors. A new method for HA analysis, employing ion exchange (IEX) chromatography to fractionate HA by size and specific quantification of each size fraction by competitive enzyme-linked sorbent assay (ELSA), was developed. When separated into four fractions, milk HA with M ≤ 20 kDa, M ∼ 20 to 60 kDa, and M ∼ 60 to 110 kDa comprised averages of 1.5, 1.4, and 2.0% of the total HA, respectively. The remaining 95% was HA with M ≥ 110 kDa. Electrophoretic analysis of the higher M HA from 13 samples showed nearly identical M distributions, with an average M of approximately 440 kDa. This higher M HA component in human milk is proposed to bind to CD44 and to enhance human beta defensin 2 (HBD2) induction by the low-M HA components.
KW - Electrophoresis
KW - Hyaluronan
KW - Ion exchange
KW - Milk
KW - Molecular mass
KW - Quantification
UR - http://www.scopus.com/inward/record.url?scp=84923339196&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84923339196&partnerID=8YFLogxK
U2 - 10.1016/j.ab.2014.12.020
DO - 10.1016/j.ab.2014.12.020
M3 - Article
C2 - 25579786
AN - SCOPUS:84923339196
VL - 474
SP - 78
EP - 88
JO - Analytical Biochemistry
JF - Analytical Biochemistry
SN - 0003-2697
ER -