Implementation of a protein profiling platform developed as an academic-pharmaceutical industry collaborative effort.
As much attention has devoted to the proteome research during the last few years, biomarker discovery has become an increasingly hot area, potentially enabling the development of new assays for diagnosis and prognosis of severe diseases. This is the field of research interest where efforts originating from both academic and industrial groups should jointly work on solutions. In this paper, we would like to demonstrate the fruitful combination of both research domains where the scientific crossroads sprout fresh ideas from the basic research domain and how these are refined and tethered to industrial standards. We will present an approach that is based on novel microfluidic devices, utilizing their benefits in processing small-volume samples. Our biomarker discovery strategy, built around this platform, involves optimized samples processing (based on SPE and sample enrichment) and fast MALDI-MS readout. The identification of novel biomarkers at low-abundance level has been achieved by the utilization of a miniaturized sample handling platform, which offers clean-up and enrichment of proteins in one step. Complete automation has been realized in the form of a unique robotic instrumentation that is able to extract and transfer 96 samples onto standard MALDI target plates with high throughput. The developed platform was operated with a 60 sample turnaround per hour allowing sensitivities in femtomol regions of medium- and low-abundant target proteins from clinical studies on samples of multiple sclerosis and gastroesophageal reflux disease. Several proteins have been identified as new biomarkers from cerebrospinal fluid and esophagus epithelial cells.
Végvári
Akos
A
Department of Electrical Measurements, Lund University, Lund, Sweden.
Magnusson
Mattias
M
Wallman
Lars
L
Ekström
Simon
S
Bolmsjö
Gunnar
G
Nilsson
Johan
J
Miliotis
Tasso
T
Ostling
Jörgen
J
Kjellström
Sven
S
Ottervald
Jan
J
Franzén
Bo
B
Hultberg
Hans
H
Marko-Varga
György
G
Laurell
Thomas
T
D000047:Academies and Institutes; D015415:Biomarkers / Q000378:metabolism*; D035843:Biomedical Research / Q000458:organization & administration; D003299:Cooperative Behavior; D004345:Drug Industry; D015180:Electrophoresis, Gel, Two-Dimensional; D004847:Epithelial Cells / Q000378:metabolism; D004947:Esophagus / Q000378:metabolism; D005764:Gastroesophageal Reflux / Q000378:metabolism; D006801:Humans; D046210:Microfluidic Analytical Techniques; D009103:Multiple Sclerosis / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D012371:Robotics; D052616:Solid Phase Extraction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D015415:Biomarkers; D020543:Proteome