Academic Journal
Rapid microfluidic sample preparation for mass spectrometric analysis of wild-type and mutant BRAF protein.
| Τίτλος: | Rapid microfluidic sample preparation for mass spectrometric analysis of wild-type and mutant BRAF protein. |
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| Συγγραφείς: | Lin YH; Department of Biomedical Engineering, Chang Gung University, Taoyuan City, Taiwan; Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan City, Taiwan. Electronic address: yenheng@mail.cgu.edu.tw., Chang HY; Department of Biomedical Engineering, Chang Gung University, Taoyuan City, Taiwan., Wu CC; Molecular Medicine Research Center, Chang Gung University, Taoyuan City, Taiwan., Hsiao YC; Molecular Medicine Research Center, Chang Gung University, Taoyuan City, Taiwan; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan City, Taiwan; Department of Otolaryngology-Head and Neck Surgery, Chang Gung Memorial Hospital at Linkou, Taoyuan City, Taiwan., Wen YH; Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan City, Taiwan; School of Medicine, National Tsing Hua University, Hsinchu City, Taiwan; School of Medicine, Chang Gung University, Taoyuan City, Taiwan., Yu JS; Molecular Medicine Research Center, Chang Gung University, Taoyuan City, Taiwan; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan City, Taiwan; Department of Otolaryngology-Head and Neck Surgery, Chang Gung Memorial Hospital at Linkou, Taoyuan City, Taiwan; Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, 33302, Taoyuan, Taiwan. Electronic address: yusong@mail.cgu.edu.tw. |
| Πηγή: | Analytica chimica acta [Anal Chim Acta] 2026 Mar 15; Vol. 1391, pp. 345146. Date of Electronic Publication: 2026 Jan 23. |
| Τύπος έκδοσης: | Journal Article; Research Support, Non-U.S. Gov't |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 0370534 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-4324 (Electronic) Linking ISSN: 00032670 NLM ISO Abbreviation: Anal Chim Acta Subsets: MEDLINE |
| Imprint Name(s): | Publication: Amsterdam : Elsevier Original Publication: Amsterdam. |
| Ιατρικοί όροι (MeSH): | Proto-Oncogene Proteins B-raf*/genetics , Proto-Oncogene Proteins B-raf*/analysis , Microfluidic Analytical Techniques*/instrumentation , Mutant Proteins*/analysis , Mutant Proteins*/genetics, Humans ; Mass Spectrometry ; Mutation |
| Περίληψη: | Background: The BRAF V600E mutant protein is a valuable biomarker for the diagnosis and prognosis of colorectal cancer. Quantitative detection by mass spectrometry (MS) requires purification from complex cell extracts and digestion into surrogate peptides, a process that traditionally takes at least ∼12 h, which is time-consuming and labor-intensive, limiting clinical applicability. Results: We present an automated microfluidic sample-preparation chip that integrates immunoprecipitation, multistep washing, and on-bead tryptic digestion by integrating pneumatically driven micromixers, microvalves, and magnetically guided beads. On-bead digestion was employed to eliminate multiple buffer-exchange steps, simplifying fluidic control and producing more BRAF peptides than the conventional elution-digestion workflow, in which only ∼50 % of captured proteins are recovered. Starting from clarified cell lysate, the device produces peptide samples within approximately 2.5-3 h. Significance and Novelty: Although individual components of the workflow have been reported previously, the present platform uniquely achieves end-to-end integration and automation of immuno-MS sample preparation. This work emphasizes operational simplicity and rapid turnaround time, providing a practical solution for MS-based mutant protein analysis in translational and precision medicine applications. (Copyright © 2026 Elsevier B.V. All rights reserved.) |
| Competing Interests: | Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. |
| Contributed Indexing: | Keywords: Mass spectrometry; Microfluidics; Mutant BRAF; Proteomics; Sample preparation |
| Substance Nomenclature: | EC 2.7.11.1 (Proto-Oncogene Proteins B-raf) EC 2.7.11.1 (BRAF protein, human) 0 (Mutant Proteins) |
| Entry Date(s): | Date Created: 20260209 Date Completed: 20260707 Latest Revision: 20260806 |
| Update Code: | 20260806 |
| DOI: | 10.1016/j.aca.2026.345146 |
| PMID: | 41663222 |
| Βάση Δεδομένων: | MEDLINE |
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