Protein identification from dried nipple aspirate fluid on Guthrie cards using mass spectrometry

Delmonico,Lucas; Areias,Vivian  Rabello; Pinto,Rodrigo  César; Matos,Cintia  Da Silva; Rosa,Marco  Felipe Franco; De Azevedo,Carolina  Maria; Alves,Gilda

doi:10.3892/mmr.2015.3432

Protein identification from dried nipple aspirate fluid on Guthrie cards using mass spectrometry

Authors:
- Lucas Delmonico
- Vivian Rabello Areias
- Rodrigo César Pinto
- Cintia Da Silva Matos
- Marco Felipe Franco Rosa
- Carolina Maria De Azevedo
- Gilda Alves
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Affiliations: Applied Genetics Laboratory, Haematology Service, National Cancer Institute, Rio de Janeiro, 20230-130, Brazil, Radiology Service, Gaffrée e Guinle University Hospital, Rio de Janeiro, 20270-004, Brazil
Published online on: March 5, 2015 https://doi.org/10.3892/mmr.2015.3432
Pages: 159-164
Copyright: © Delmonico et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Nipple aspirate fluid (NAF) requires investigation as a potential source of biomarkers for early diagnosis or risk assessment in breast cancer and other breast disorders. The present study demonstrated that proteins were easily extracted from dried NAF spots on Guthrie cards and were suitable for mass spectrometry analysis. NAF was obtained from 80 women, collected on Guthrie cards, between 2007 and 2010. The NAF‑proteins were extracted from the card by incubating the card in water. These proteins were then quantified and separated using one‑dimensional, 12% SDS‑PAGE, gel electrophoresis and on high‑resolution gradient gels at different concentrations (4‑12, 8‑16 and 4‑20%). The bands with the most abundant proteins were excised from the gradient gels and the proteins were identified by liquid chromatography quadrupole time of flight. Immunoglobulins, Zn‑α2‑glicoprotein, apoliprotein D and prolactin inducible protein were among those identified. The NAF-Guthrie card collection method has not been applied previously, however, NAF proteins have been identified using other collecting techniques, confirming the feasibility of the NAF Guthrie card collection method for analyzing the proteins within NAF. The NAF‑Guthrie card collecting method has multiple advantages, including being inexpensive, non‑invasive, reliable and painless, and the cards can be stored at room temperature. Examining NAF may assist in identifying individuals at a higher risk of breast cancer and in improving patient prognosis.

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