Ultrasonic Enhanced Applications in Proteomics Workflows: single probe versus multiprobe

DOI: 10.5584/jiomics.v1i1.55

Authors

  • Luz Fernandes REQUIMTE, Departamento de Química, Centro de Química Fina e Biotecnologia,Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2829-516 Monte de Caparica, Portugal http://orcid.org/0000-0003-0457-1575
  • Hugo M. Santos REQUIMTE, Departamento de Química, Centro de Química Fina e Biotecnologia,Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2829-516 Monte de Caparica, Portugal https://orcid.org/0000-0002-6032-8679
  • J. D. Nunes-Miranda Physical Chemistry Department. Science Faculty, University of Vigo. E- 32004. Ourense Spain
  • Carlos Lodeiro REQUIMTE, Departamento de Química, Centro de Química Fina e Biotecnologia,Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2829-516 Monte de Caparica, Portugal https://orcid.org/0000-0001-5582-5446
  • José L. Capelo REQUIMTE, Departamento de Química, Centro de Química Fina e Biotecnologia,Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2829-516 Monte de Caparica, Portugal https://orcid.org/0000-0001-6276-8507

Abstract

A 96-well plate-based platform in conjunction with an ultrasonic multiprobe of four tips was assessed to develop various fast proteomics workflows for gel-based proteomics. The use of such protocols reduce sample time and handling, allowing rapid processing whilst reducing the risk of contamination. The procedure reduces the time to indentify proteins separated by gel electrophoresis to just 8 min/each. In addition, the ultrasonic multiprobe was compared with the single probe as a tool to obtain high sample throughput in proteomics workflows entailing identification and/or quantification of proteins using mass-spectrometry based approaches. The 18-O labeling-based method was used to study the type of peptides extracted from the gels when the extraction was done with the aid of ultrasonic energy. The assessment was done in ten standard proteins separated by gel elecrophoresis. Two proteins obtained from D. desulfuricans, and from Cyprinus carpio,  Split-Soret cytochrome c, and Vitellogenin respectively, were also indentified as a further proof-of-the concept.

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Published

2021-02-14