Proteomics, the large-scale study of proteins, has revolutionized our understanding of biological processes and disease mechanisms. Enhanced multiplexing technology has significantly advanced the field by allowing simultaneous analysis of multiple samples, thereby increasing throughput, precision, and quantitative accuracy.
Cutting-Edge Multiplexing Technologies
In the realm of proteomics, multiplexing technologies have revolutionized the ability to analyze multiple samples simultaneously, significantly enhancing throughput, precision, and depth of analysis, including [1]:
Tandem Mass Tag (TMT) Technology
Tandem Mass Tag (TMT) technology utilizes isobaric chemical tags to label peptides from different samples, allowing them to be indistinguishably analyzed during liquid chromatography but producing distinct reporter ions during mass spectrometry. This method is renowned for its ability to simultaneously analyze up to 16 samples, significantly increasing throughput and data acquisition efficiency.
Isobaric Tags for Relative and Absolute Quantitation (iTRAQ)
iTRAQ is another widely used multiplexing technology that labels the N-terminus and side-chain amines of peptides with isobaric tags. These tags produce identical mass signals during chromatography but generate unique reporter ions during fragmentation in the mass spectrometer. This technology supports the simultaneous analysis of up to 8 samples, offering high sensitivity and quantitative accuracy.
Hyperplexing with TMTpro
TMTpro represents an advancement over traditional TMT technology, allowing for even higher multiplexing capabilities. By utilizing a more complex isotopic labeling strategy, TMTpro can analyze up to 18 samples concurrently. This increase in multiplexing capacity is achieved without compromising the resolution or quantitative accuracy of the analysis.
Data-Independent Acquisition (DIA)
Data-Independent Acquisition (DIA) is a mass spectrometry technique that differs from traditional Data-Dependent Acquisition (DDA) by simultaneously fragmenting all ions within a selected mass range, rather than selecting specific precursor ions. This method provides a comprehensive and unbiased acquisition of proteomic data, making it highly suitable for quantitative studies.
WATH-MS (Sequential Window Acquisition of All Theoretical Mass Spectra)
SWATH-MS is a form of DIA that involves sequentially isolating precursor ions in defined m/z windows, fragmenting them, and recording the resulting fragment ions. This method combines the comprehensive data acquisition of DIA with the quantitative accuracy of targeted proteomics, making it a powerful tool for high-throughput proteomics.
Applications in Disease Research
Typical applications of these advanced multiplexing techniques in various fields of biomedical research are listed below:
Alzheimer's Disease
Enhanced multiplexing technology has been pivotal in studying neurodegenerative diseases like Alzheimer's. Deep multilayer brain proteomics using TMT has identified critical molecular networks involved in disease progression. For instance, Bai et al. [2] utilized multiplexed proteomics to analyze post-mortem brain tissues, revealing dysregulated pathways and potential therapeutic targets.
Cancer Research
Multiplexing has also significantly impacted cancer research. Proteogenomic studies combining proteomics and genomics have provided a comprehensive view of cancer biology. Dou et al. [3] used TMT-based proteomics to characterize endometrial carcinoma, uncovering novel biomarkers and therapeutic targets.
Aging and Metabolism
Multiplexed proteomics is invaluable in aging and metabolic studies. Yu et al. [4] used TMT to analyze protein changes in aging mice, identifying pathways involved in aging and potential interventions. Similarly, proteomic studies in metabolic diseases have revealed critical insights into the molecular mechanisms underlying conditions like diabetes and obesity.
Cardiovascular Diseases
Proteomics has advanced the understanding of cardiovascular diseases. Raafs et al. [5] utilized multiplexing to identify sex-specific biomarkers predicting heart failure, providing new avenues for personalized treatment strategies. Such studies underscore the importance of proteomics in uncovering the molecular basis of complex diseases.
Enhanced multiplexing technology has transformed proteomics, enabling high-throughput, precise, and comprehensive protein analysis. Through techniques like TMT and iTRAQ, and powerful tools like advanced mass spectrometry and sophisticated data analysis software, researchers can now delve deeper into the proteome. Our company is a leading supplier of proteomics services. Contact us to learn more about how we can support your scientific endeavors and help you achieve your goals.
References
- Bowser BL, Robinson RAS. Enhanced Multiplexing Technology for Proteomics. Annu Rev Anal Chem (Palo Alto Calif). 2023 Jun 14;16(1):379-400.
- Bai B, Wang X, Li Y, et al. Deep Multilayer Brain Proteomics Identifies Molecular Networks in Alzheimer's Disease Progression. Neuron. 2020 Mar 18;105(6):975-991.e7.
- Dou Y, Kawaler EA, Cui Zhou D, et al. Proteogenomic Characterization of Endometrial Carcinoma. Cell. 2020 Feb 20;180(4):729-748.e26.
- Yu Q, Xiao H, Jedrychowski MP, et al. Sample multiplexing for targeted pathway proteomics in aging mice. Proc Natl Acad Sci U S A. 2020 May 5;117(18):9723-9732.
- Raafs A, Verdonschot J, Ferreira JP, et al. Identification of sex-specific biomarkers predicting new-onset heart failure. ESC Heart Fail. 2021 Oct;8(5):3512-3520.
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