Ultrasensitive Detection of Cardiac Troponin I via CRISPR/Cas12a-Mediated Liposomal Amplification Coupled with Electrospray Ionization Mass Spectrometry

  • Anal Chem. 2026 Jan 27;98(3):2183-2190. doi: 10.1021/acs.analchem.5c05804.
Xiang Chen  1  2 ,  Chunxiang Mao  3 ,  Yuanji Gao  4 ,  Chaoting Shi  1 ,  Yu Wang  1 ,  Zhuolin Jin  1  2 ,  Bing Xia  1 ,  Yan Zhou  1
Affiliations
  • 1. Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan 610213, People's Republic of China.
  • 2. University of Chinese Academy of Sciences, Beijing 101408, People's Republic of China.
  • 3. Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 611137, People's Republic of China.
  • 4. College of Chemistry and Materials Science, Sichuan Normal University, Chengdu, Sichuan 610068, People's Republic of China.
PMID: 41533833 DOI: 10.1021/acs.analchem.5c05804
Abstract

Direct quantitative analysis of low-abundance protein biomarkers by electrospray ionization mass spectrometry (ESI-MS) remains challenging due to poor ionization efficiency and matrix interferences. Herein, we report an ultrasensitive analytical platform, termed CRISPR/Cas12a-mediated liposomal amplification coupled with electrospray ionization mass spectrometry (CMLA-MS), that overcomes this limitation by integrating CRISPR/Cas12a-mediated dual-cascade signal amplification with an ESI-MS readout. The strategy converts the detection of poorly ionizable protein molecules into the quantification of numerous, highly ionizable small-molecule reporters: proteins trigger Cas12a trans-cleavage (first amplification), which subsequently cleaves single-stranded DNA (ssDNA) probes anchored to signal-loaded liposomes, causing the burst release of thousands of MS-detectable reporters (second, physical amplification). This dual-amplification strategy enabled an exceptionally low limit of detection (LOD) of 10.8 fg/mL, and the method successfully quantified cardiac troponin I (cTnI) in clinical serum samples with high recoveries (90.3-101.6%).

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