title: Development of high-sensitive ELISA method for detection of adipophilin levels in human colostrum and breast milk.
abstract: OBJECTIVE
  To develop and validate high-sensitive (hs) ELISA method for detection of adipophilin (adipose differentiation-related protein, ADRP) in human breast milk (BM) and to analyze adipophilin levels in BM during 12 months of lactation.
  
  
  METHODS
  ADRP levels were determined using hsELISA method (Biovendor-Laboratory Medicine, Inc.) in colostrum (D0) and BM of 72 mothers was collected 1, 3, 6, and 12 months following delivery (M1, 3, 6, 12).
  
  
  RESULTS
  ADRP was detectable in BM up to 12 months of lactation. Mean levels at D0 were 1.98 ± 0.12; M1, 2.83 ± 0.21; M3, 2.39 ± 0.17; M6, 2.57 ± 0.16; and at M12 3.25 ± 0.21 μg/ml. Significantly higher levels of ADRP were found in M1 and M12 when compared to D0 and in M12 when compared to M3 (overall P = 0.0001). No significant correlation was seen between ADRP levels in BM and adiponectin, body weight of infants, their birth length, body weight gain during the first year of life, or BMI of mothers before pregnancy.
  
  
  CONCLUSIONS
  We developed and validated hsELISA for detection of ADRP in human BM. ADRP was detectable in human BM during the whole 12 months of lactation period and its levels were intraindividually well-conserved.
authors.affiliation: Department of Paediatrics, 2nd Faculty of Medicine and University Hospital Motol, Charles University in Prague, Prague, Czech Republic.
authors.forename: Katarina
authors.initials: K
authors.lastname: Mitrova
authors.forename: Michal
authors.initials: M
authors.lastname: Karpisek
authors.forename: Marianna
authors.initials: M
authors.lastname: Durilova
authors.forename: Lavinia Gabriela
authors.initials: LG
authors.lastname: Dragusin
authors.forename: Jiri
authors.initials: J
authors.lastname: Nevoral
authors.forename: Jiri
authors.initials: J
authors.lastname: Bronsky
mesh_terms: D003126:Colostrum / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D007774:Lactation; D008565:Membrane Proteins / Q000378:metabolism*; D008895:Milk, Human / Q000378:metabolism*; D000070780:Perilipin-2; D011247:Pregnancy; D013997:Time Factors
chemical_list: D008565:Membrane Proteins; C000602982:PLIN2 protein, human; D000070780:Perilipin-2
keywords: ADRP; ELISA; adipophilin; human breast milk; nutritional programming