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Portable Biosensor for Label-Free Protein Detection
KTH, School of Engineering Sciences (SCI), Applied Physics.
2025 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesisAlternative title
Biosensor för proteindetektering (Swedish)
Abstract [en]

Streaming current–based biosensing provides a label-free and highly sensitive method for detecting molecular interactions at solid–liquid interfaces. In this work, a microfluidic platform integrating an Elveflow flow controller and Keithley 2636A picoammeter was developed and validated for quantitative biosensing of biotin-binding proteins. A data-analysis pipeline was established, including signal synchronization, baseline correction, and plateau detection, complemented by theoretical modeling of flow–pressure relations and rigorous noise analysis.To enable uniform comparison across channels, the operating plateau's RMS noise was used to establish the minimum detectable signal (MDS). Although Streptavidin, NeutrAvidin, and Avidin showed concentration-dependent responses, the absolute streaming current values were very high, most likely as a result of background noise. Protein-dependent changes in streaming current that were compatible with their net charge were qualitatively validated by the trends that were observed. Non-specific binding was verified to be suppressed by negative control validation, with very slight sub-threshold residuals resulting from fluidic or instrumental variability.The results establish a reliable framework for streaming current biosensing, demonstrating both high sensitivity and robustness against noise and artifacts. The presented platform provides a solid foundation for extending label-free biosensing to more complex analytes, real sample matrices, and multiplexed detection schemes.

Abstract [sv]

Biosensorteknik baserad på strömningsström erbjuder en etikettfri och mycket känslig metod för att detektera molekylära interaktioner vid fasta–vätskegränssnitt. I detta arbete utvecklades och validerades en mikrofluidisk plattform som integrerar en Elveflow-flödeskontroller och en Keithley 2636A picoammeter för kvantitativ biosensorik av biotinbindande proteiner. En dataanalytisk pipeline etablerades, innefattande signalsynkronisering, baslinjekorrigering och platådetektion, kompletterad med teoretisk modellering av flödes–tryckrelationer samt rigorös brusanalys.För att möjliggöra en enhetlig jämförelse mellan kanaler användes driftplatåns RMS-brus för att fastställa den minsta detekterbara signalen (MDS). Även om Streptavidin, NeutrAvidin och Avidin uppvisade koncentrationsberoende svar, var de absoluta strömningsströmsvärdena mycket höga, sannolikt som ett resultat av bakgrundsbrus. Proteinspecifika förändringar i strömningsström som överensstämde med deras nettoladdning validerades kvalitativt genom de observerade trenderna. Ospecifik bindning verifierades vara undertryckt genom negativ kontrollvalidering, med mycket små subtröskelrester som härrör från flödes- eller instrumentvariabilitet.Resultaten etablerar en tillförlitlig ram för biosensorik baserad på strömningsström och visar både hög känslighet och robusthet mot brus och artefakter. Den presenterade plattformen utgör en solid grund för att utvidga etikettfri biosensorik till mer komplexa analyter, verkliga provmatriser och multiplexade detektionsscheman.

Place, publisher, year, edition, pages
2025.
Series
TRITA-SCI-GRU ; 2025:456
Keywords [en]
Streaming current biosensing, Label-free detection, Microfluidics, Streptavidin, NeutrAvidin, Avidin, Minimum Detectable Signal (MDS)
Keywords [sv]
Strömningsströmsbiosensorik, Etikettfri detektion, Mikrofluidik, Streptavidin, NeutrAvidin, Avidin, Minsta detekterbara signal (MDS)
National Category
Physical Sciences
Identifiers
URN: urn:nbn:se:kth:diva-371238OAI: oai:DiVA.org:kth-371238DiVA, id: diva2:2004295
External cooperation
Uppsala University
Subject / course
Physics
Educational program
Master of Science - Engineering Physics
Supervisors
Examiners
Available from: 2025-10-07 Created: 2025-10-07 Last updated: 2025-10-07Bibliographically approved

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