Ultrasonic-based membrane aided sample preparation of urine proteomes

Author(s)

Jesus, Jemmyson Romario ^{[1, 2, 3]} ; Santos, Hugo M. ^{[4, 2]} ; Lopez-Fernandez, H. ^{[2, 5, 6]} ; Lodeiro, Carlos ^{[4, 2]} ; Zezzi Arruda, Marco Aurelio ^{[1, 3]} ; Capelo, J. L. ^{[4, 2]}

Affiliation

^[1] Univ Campinas Unicamp, Inst Chem, Natl Inst Sci & Technol Bioanalyt, POB 6154, BR-13083970 Campinas, SP - Brazil

^[2] Univ Nova Lisboa, Fac Sci & Technol, Chem Dept, UCIBIO REQUIMTE, P-2829516 Caparica - Portugal

^[3] Univ Campinas UNICAMP, Inst Chem, Spectrometry Sample Preparat & Mechanizast Grp GE, POB 6154, BR-13083970 Campinas, SP - Brazil

^[4] ProteoMass Sci Soc, Madan Parque, Rua Inventores, P-2825182 Caparica - Portugal

^[5] Univ Vigo, Escuela Super Ingn Informat, Edificio Politecn, Lagoas S-N, Orense 32004 - Spain

^[6] Ctr Singular Invest Galicia, Ctr Invest Biomed, Vigo - Spain

• Document type: Journal article
• Source: Talanta; v. 178, p. 864-869, FEB 1 2018.
• Web of Science Citations: 3

Abstract

A new ultrafast ultrasonic-based method for shotgun proteomics as well as label-free protein quantification in urine samples is developed. The method first separates the urine proteins using nitrocellulose-based membranes and then proteins are in-membrane digested using trypsin. The enzymatic digestion process is accelerated from overnight to four minutes using a sonoreactor ultrasonic device. Overall, the sample treatment pipeline comprising protein separation, digestion and identification is done in just 3 h. The process is assessed using urine of healthy volunteers. The method shows that male can be differentiated from female using the protein content of urine in a fast, easy and straightforward way. 232 and 226 proteins are identified in urine of male and female, respectively. From this, 162 are common to both genders, whilst 70 are unique to male and 64 to female. From the 162 common proteins, 13 are present at levels statistically different (p < 0.05). The method matches the analytical minimalism concept as outlined by Halls, as each stage of this analysis is evaluated to minimize the time, cost, sample requirement, reagent consumption, energy requirements and production of waste products. (AU)

• FAPESP's process: 16/07384-7 - Elementary bioimaging through laser ablation and inductively coupled plasma mass spectrometry
• Grantee: Marco Aurelio Zezzi Arruda
• Support Opportunities: Regular Research Grants