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Utilization of chromogenic enzyme substrates for signal amplification in multiplexed detection of biomolecules using surface mass spectrometry

 Hee-Kyung Na  ;  Hyun Kyong Shon  ;  Hye Young Son  ;  Eunji Jang  ;  Sunho Joh  ;  Yong-Min Huh  ;  David G Castner  ;  Tae Geol Lee 
 SENSORS AND ACTUATORS B-CHEMICAL, Vol.332 : 129452, 2021-04 
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Enzyme-based signal amplification in mass spectrometry ; ToF-SIMS ; miRNA sensing ; molecular signal enhancer ; single base mismatch discrimination
MicroRNAs (miRNAs) are important post-transcriptional gene regulators and can serve as potential biomarkers for many diseases. Most of the current miRNA detection techniques require purification from biological samples, amplification, labeling, or tagging, which makes quantitative analysis of clinically relevant samples challenging. Here we present a new strategy for the detection of miRNAs with uniformity over a large area based on signal amplification using enzymatic reactions and measurements using time-of-flight secondary ion mass spectrometry (ToF-SIMS), a sensitive surface analysis tool. This technique has high sequence specificity through hybridization with a hairpin DNA probe and allows the identification of single-base mismatches that are difficult to distinguish by conventional mass spectrometry. We successfully detected target miRNAs in biological samples without purification, amplification, or labeling of target molecules. In addition, by adopting a well-known chromogenic enzymatic reaction from the field of biotechnology, we extended the use of enzyme-amplified signal enhancement ToF (EASE-ToF) to protein detection. Our strategy has advantages with respect to scope, quantification, and throughput over the currently available methods, and is amenable to multiplexing based on the outstanding molecular specificity of mass spectrometry (MS). Therefore, our technique not only has the potential for use in clinical diagnosis, but also provides evidence that MS can serve as a useful readout for biosensing to perform multiplexed analysis extending beyond the limitations of existing technology.
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1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Radiology (영상의학교실) > 1. Journal Papers
Yonsei Authors
Son, Hye Yeong(손혜영) ORCID logo https://orcid.org/0000-0001-5977-6784
Huh, Yong Min(허용민) ORCID logo https://orcid.org/0000-0002-9831-4475
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