Optimisation and comparison of orthogonal methods for separation and characterisation of extracellular vesicles to investigate how representative infant milk formula is of milk

Anindya Mukhopadhya, Jessie Santoro, Barry Moran, Zivile Useckaite, Lorraine O’Driscoll

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Many infants are fed infant milk formula (IMF). However, IMF production from skim milk (SM) involves harsh treatment. So, we hypothesised that the quantity and/or quality of extracellular vesicles (EVs) in IMF may be reduced. Thus, firstly, we aimed to optimise separation of EVs from IMF and SM and, secondly, we aimed to compare the EV isolates from these two sources. Prior to EV isolation, abundant casein micelles of similar sizes to EVs were removed by treating milk samples with either acetic acid or hydrochloric acid. Samples progressed to differential ultracentrifugation (DUC) or gradient ultracentrifugation (GUC). EV characterisation included BCA, SDS-PAGE, nanoparticle tracking (NTA), electron microscopy (TEM), immunoblotting, and imaging flow cytometry (IFCM). Reduced EV concentrations were found in IMF. SM-derived EVs were intact, while IMF contained disrupted EV-like structures. EV biomarkers were more abundant with isolates from SM, indicating EV proteins in IMF are compromised. Altogether, a suitable method combining acid pre-treatment with GUC for EV separation from milk products was developed. EVs appear to be substantially compromised in IMF compared to SM.

Original languageEnglish
Article number129309
Number of pages11
JournalFood Chemistry
Volume353
Early online date23 Feb 2021
DOIs
Publication statusPublished - 15 Aug 2021
Externally publishedYes

Keywords

  • Extracellular vesicles (EVs)
  • milk
  • infant milk formula (IMF)
  • pre-treatment
  • developed methodologies
  • EV-TRACKED
  • Developed methodologies
  • Pre-treatment
  • Infant milk formula (IMF)
  • Milk

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