RESUMEN
The analysis of protein antigens as biomarkers in clinical samples is particularly helpful for the early diagnosis of diseases. However, this is difficult to accomplish owing to the presence of the antigens in trace amounts as well as the complexity of the matrixes in clinical samples. In this study, a lab-on-membrane platform that can be combined with paper spray ionization mass spectrometry was developed for the in situ high-throughput sensitive detection of the prostate-specific antigen (PSA). The sensitivity of the proposed platform was enhanced via two strategies: (1) the synthesis of a biotin-streptavidin scaffold caused an increase in the capturing efficiency of PSA by a factor of 5 and (2) the immobilization of a large number of mass tag molecules on the gold nanoparticles allowed for the amplification of the mass spectrometry signals. The limit of detection was approximately 3.0 pg mL-1. The selectivity to PSA was guaranteed by using an antibody-aptamer pairing sandwich immunoassay, and PSA detection was unaffected even when other protein antigens (carcinoembryonic antigen and carbohydrate antigen 125) were present. The modified membranes maintained their performance for at least 30 days when stored at 4 °C. Finally, analysis of human serum samples confirmed that the PSA concentration as determined using the proposed platform was consistent with that determined with a conventional chemiluminescent immunoassay. Thus, this PSA analyzing platform is suitable for prostate cancer diagnosis in clinical settings.
Asunto(s)
Técnicas Biosensibles , Papel , Antígeno Prostático Específico/sangre , Oro/química , Humanos , Nanopartículas del Metal/química , Sondas Moleculares/química , Estructura Molecular , Glicoles de Propileno/química , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Comparative studies of molecular alterations upon cancer between mice and humans are of great importance in order to determine the relevance of research involving mouse cancer models to the development of diagnostic and therapeutic approaches in clinical practice as well as for the mechanistic studies of pathology in humans. Herein, using molecular fingerprinting by internal extractive electrospray ionization mass spectrometry (iEESI-MS), we identified 50 differential signals in mouse liver tissue and 62 differential signals in human liver tissue that undergo significant intensity alterations (variable importance in the project (VIP) >1.0) upon liver cancer, out of which only 27 were common in both mouse and human tissues. Out of the 27 common differential signals, six types of phospholipids were also identified to undergo significant alterations in human serum upon liver cancer, including PC(34:2), PC(36:4), PC(38:6), PC(36:2), PC(38:4) and PC(42:9). Statistical analysis of the relative intensity distribution of these six identified phospholipids in serum allowed confident determination of liver cancer in humans (sensitivity 91.0%, specificity 88.0%, and accuracy 90.0%). Our results indicate that, despite the significant difference in the overall alterations of phospholipid profiles upon liver cancer between humans and mice, the six identified 'core' differential phospholipids of liver cancer found in the liver tissues of both humans and mice as well as in human serum show high potential as a minimal panel for the rapid targeted diagnosis of liver cancer with high accuracy, sensitivity and specificity using direct mass spectrometry (MS) analysis.
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Neoplasias Hepáticas , Fosfolípidos , Animales , Humanos , Ratones , Sensibilidad y Especificidad , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Rapid quantitative determination of bulk molecular concentration in solid samples without sample pretreatment is demonstrated using the internal extractive electrospray ionization-mass spectrometry (iEESI-MS) analysis of six ß-agonists, including salbutamol (Sal), clenbuterol (Cle), ractopamine (Rac), terbutaline (Ter), tulobuterol (Tul), brombuterol (Bro), in pork tissue samples. Single sample analysis only required 1 min. The linear range of detection was about 0.01-1000 µg/kg (R2 > 0.9994). The limit-of-detection (LOD) varied from 0.002 µg/kg for Sal to 0.006 µg/kg for Tul. Relative standard deviation (RSD) of quantitation was in the range 6.5-11.3%. The analytical results were validated by gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography-mass spectrometry (LC-MS), showing the accuracy rates of 92-105%. The current study extends the power of ambient MS as a method for the quantification of molecules at the surface of solid samples (e.g., in µg/cm2 units) toward the quantification of molecules in bulk sample volume (i.e., in µg/kg units), which is commonly required in food safety control, biomedical analysis, public security, and many other disciplines.
Asunto(s)
Agonistas de Receptores Adrenérgicos beta 2/análisis , Contaminación de Alimentos/análisis , Carne Roja/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos , Animales , Límite de Detección , PorcinosRESUMEN
Investigation on the distribution of persistent organic pollutants (POPs) in aquatic organisms is of great importance for exploring the biological toxicity and health risks of environmental pollutants. In this study, a liquid extraction surface analysis mass spectrometry (LESA-MS) method was developed for rapid and in situ analysis of the spatial distribution of perfluoroalkyl substances (PFASs) in zebrafish. By combining the high-precision automated moving platform of LESA device and the high-resolution MS, quantitative analysis of perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) in zebrafish tissue section were easily achieved. A tissue-specific ionization efficiency factor (TSF) strategy was also proposed to correct the matrix effect in different parts of zebrafish tissue. By using the developed method, high sensitive and efficient imaging of PFOA and PFOS in zebrafish tissue was achieved, and the distributions of PFOA and PFOS in descending order were gills, organs, roes, pelvic fin, muscle, and brain. The experimental results demonstrated that the coupling of LESA-MS method with TFS strategy is an efficient and reliable approach for monitoring the content distribution of environmental pollutants in biological tissues.
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Ácidos Alcanesulfónicos , Contaminantes Ambientales , Fluorocarburos , Contaminantes Químicos del Agua , Animales , Caprilatos , Contaminantes Ambientales/análisis , Fluorocarburos/análisis , Espectrometría de Masas en Tándem , Pez CebraRESUMEN
PURPOSE: Increased use of low-dose spiral computed tomography (LDCT: low-dose computed tomography) screening has contributed to more frequent incidental detection of peripheral lung nodules, part of them were adenocarcinoma, which need to be further evaluated to establish a definitive diagnosis. Here, our primary objective was to evaluate the ambient mass spectrometry (AMS) sputum analysis as a non-invasive lung adenocarcinoma (LAC) diagnosis solution. PATIENTS AND METHODS: Neutral desorption extractive electrospray ionization mass spectrometry (ND-EESI-MS) and collision induced dissociation (CID) were used to detect sputum metabolites from 143 spontaneous sputum samples. Partial least squares-discriminant analysis (PLS-DA) was used to refine the biomarker panel, whereas orthogonal PLS-DA (OPLS-DA) was used to operationalize the enhanced biomarker panel for diagnosis. RESULTS: In this approach, 19 altered metabolites were detected by ND-EESI-MS from 76 cases of LAC and 67 cases of control. Significance testing and receiver operating characteristic (ROC) analysis identified 5 metabolites [hydroxyphenyllactic acid, phytosphingosine, N-nonanoylglycine, sphinganine, S-carboxymethyl-L-cysteine] with p <0.05 and AUC >0.75, respectively. Evaluation of model performance for prediction of LAC resulted in a cross-validation classification accuracy of 87.9%. Metabolic pathway analysis showed that sphingolipid metabolism, fatty acid metabolism, carnitine synthesis and Warburg effect were most impacted in response to disease. CONCLUSION: This study indicates that the application of ND-EESI-MS to sputum analysis can be used as a non-invasive detection of peripheral lung nodules. The use of sputum metabolite biomarkers may aid in the development of a further evaluation program for lung adenocarcinoma.
RESUMEN
Ambient mass spectrometry imaging (AMSI) is a molecular imaging technique developed in recent years for in situ and real time visualization of the distribution of chemical compounds in biological tissues without the need of labeling or staining. With the development for more than one decade, AMSI becomes a powerful molecular imaging technique in variousfields such as forensics, metabolomics, cancer diagnosis, and drug monitoring. In this review, we describe the recent advances of AMSI for imaging biological tissues in details. Three types of AMSI techniques based on different ionization mechanisms and analytical strategies are summarized, i.e., direct desorption/ionization of analytes for AMSI, desorption and then ionization of analytes for AMSI, and extraction of analytes for AMSI, and the features of them are presented from the aspects of tissue origin, target image molecule, and spatial resolution among others. In addition, future development directions for AMSI are discussed.
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Imagen Molecular , Péptidos/análisis , Proteínas/análisis , Animales , Humanos , Espectrometría de MasasRESUMEN
Human sputum, a typical highly viscous biosample, was directly characterized at the molecular level using neutral desorption extractive electrospray ionization mass spectrometry (ND-EESI-MS) without multi-step sample pretreatment, in an attempt to provide a method for constructing the pattern recognition of rapid diagnosis of lung cancer. Under the optimal experiment conditions, glucose, amino acids, phosphoric lipids and other typical analytes in the sputum sample could be used to conduct qualitative or quantitative (in arginine) analysis. More interestingly, the full scan mass spectra from 50 patients of non-small cell lung cancer, recording the mass spectral fingerprints of sputum samples, were differentiated from the control group (50 healthy individuals) through principal component analysis (PCA). These findings suggest that valuable molecular information concealed in human sputum could be easily revealed and applied for conducting qualitative or quantitative analysis by direct ND-EESI-MS analysis.
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Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Neoplasias Pulmonares/diagnóstico , Esputo/química , Humanos , Análisis de Componente Principal , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Direct mass spectrometry analysis of metabolic effects of clenbuterol and salbutamol on pork quality at the molecular level is incredibly beneficial for food regulations, public health and the development of new anti-obesity drugs. With internal extractive electrospray ionization mass spectrometry (iEESI-MS), nutrients including creatine, amino acids, L-carnitine, vitamin B6, carnosine and phosphatidylcholines in pork tissue were identified, without sample pretreatment, using collision-induced dissociation (CID) experiments and by comparison with authentic compounds. Furthermore, normal pork samples were clearly differentiated from pork samples with clenbuterol and salbutamol via principal component analysis (PCA). Correlation analysis performed on the spectral data revealed that the above-mentioned nutrients strongly correlated with pork quality, and the absolute intensity of phosphatidylcholines in normal pork was much higher than pork contaminated by clenbuterol and salbutamol. Our findings suggested that clenbuterol and salbutamol may render effects on the activity of carnitine acyltransferase I, hence the process that L-carnitine transports long-chain fatty acids into mitochondria and the formation of phosphatidylcholines might be affected. However, the underlying metabolic mechanisms of clenbuterol and salbutamol on carnitine acyltransferase I requires more comprehensive studies in future work.