LC-MS/MS-Based Targeted Site Validation Service
Target validation analysis based on Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) is a critical method for the quantitative detection and validation of candidate drug targets or biomarkers. This technique integrates the separation capability of high-performance liquid chromatography (LC) with the high sensitivity and specificity of tandem mass spectrometry (MS/MS), enabling high-precision analysis of target proteins or small molecules in complex biological samples. Its core principle relies on targeted mass spectrometry approaches such as Selected Reaction Monitoring (SRM) or Parallel Reaction Monitoring (PRM), which precisely quantify characteristic peptides of specific targets to ensure the reliability and biological relevance of target molecules.
LC-MS/MS-based targeted site validation service is widely applied in drug discovery, disease research, personalized medicine, and biomarker validation. For instance, in drug development, this technology is used to verify whether a drug successfully acts on its intended target and to evaluate its selectivity and specificity. In precision medicine, it is employed to validate disease biomarkers, providing a basis for personalized treatment strategies. In protein interaction and signaling pathway studies, it facilitates the exploration of dynamic changes in protein-protein interactions. Additionally, this approach is utilized in biopharmaceutical development to ensure the targeting specificity and stability of protein-based drugs. With its advantages of high sensitivity, high throughput, and low background interference, LC-MS/MS has become a powerful tool in biomedical research and target validation.
Zhang, H W. et al. Journal of Proteomics, 2023.
Figure 1. Global Analysis of Protein Lysine Succinylation and Proteomic Profiles in Human Aortas.
Services at MtoZ Biolabs
Based on a high-resolution Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) platform, MtoZ Biolabs offers LC-MS/MS-based targeted site validation service with high sensitivity and specificity, ensuring data accuracy and experimental reproducibility. Our services cover sample preparation, protein extraction, enzymatic digestion, targeted peptide selection, LC-MS/MS detection, and data analysis, delivering highly reproducible results with a broad dynamic range. Utilizing targeted mass spectrometry techniques such as Selected Reaction Monitoring (SRM), Multiple Reaction Monitoring (MRM), or Parallel Reaction Monitoring (PRM), we precisely validate the expression levels of candidate targets, their post-translational modification (PTM) states, and their roles in physiological and pathological processes. This approach provides reliable data support for drug development, disease research, and personalized medicine.
Service Advantages
1. Advanced Analysis Platform
MtoZ Biolabs established an advanced LC-MS/MS-based targeted site validation service platform, guaranteeing reliable, fast, and highly accurate analysis service.
2. One-Time-Charge
Our pricing is transparent, no hidden fees or additional costs.
3. High Throughput and Broad Dynamic Range
By integrating Data-Dependent Acquisition (DDA), Data-Independent Acquisition (DIA), and targeted quantification techniques such as (PRM, MRM, SRM), we enable the simultaneous analysis of a large number of target proteins while covering a wide dynamic range. This makes our approach suitable for diverse biological systems and disease research.
4. Precise Quantification and Data Reproducibility
Utilizing stable isotope labeling strategies (SILAC, TMT/iTRAQ) or label-free quantification methods, combined with rigorous data normalization and quality control, we enhance quantification accuracy and ensure high data reproducibility, guaranteeing the reliability of experimental results.
5. Multi-Dimensional Validation and Customized Services
Our approach can be integrated with post-translational modification (PTM) analysis, protein-protein interaction studies, and multi-omics data integration. We provide customized target validation solutions tailored to specific signaling pathways or disease models, supporting precision medicine, drug development, and biomarker discovery.
Applications
1. Disease Biomarker Validation
Utilizing LC-MS/MS technology, we provide precise identification and quantification of candidate biomarkers. This approach is widely applied in early diagnosis, disease classification, and prognosis evaluation of cancer, neurodegenerative diseases, and cardiovascular disorders, driving advancements in precision medicine.
2. Drug Target Confirmation and Mechanism of Action Studies
In drug development, LC-MS/MS-based targeted site validation service verifies drug-target interactions, analyzes target protein expression levels, assesses post-translational modifications (PTMs), and monitors signaling pathway alterations. This provides critical data for drug screening and mechanism elucidation.
3. Protein Interaction and Signaling Pathway Studies
Through targeted quantification techniques such as PRM, MRM, and SRM, we analyze protein-protein interactions and dynamic changes in cellular signaling pathways. This helps researchers uncover regulatory mechanisms underlying various biological processes.
4. Biopharmaceutical Development and Quality Control
In the development of antibody drugs, protein therapeutics, and vaccines, LC-MS/MS-based targeted site validation service is used for key target verification, protein expression monitoring, and quality control, ensuring the stability and consistency of biopharmaceutical products.
5. Multi-Omics Integration Analysis
By integrating transcriptomics, metabolomics, phosphoproteomics, and other multi-omics data, LC-MS/MS target validation enables in-depth exploration of molecular regulatory networks, advancing systems biology and personalized medicine research.
Case Study
1. Direct Target Site Identification of a Sulfonyl–Triazole Covalent Kinase Probe by LC-MS Chemical Proteomics
The study aims to directly identify the targets of a sulfonyl-triazole covalent kinase probe using LC-MS-based chemical proteomics to elucidate its mechanism of action in cells. The research was conducted on human cell lines, employing a chemical proteomics approach combined with a covalent labeling strategy to identify the direct binding targets of the kinase probe. The experimental process involved the chemical synthesis of the sulfonyl-triazole probe, followed by live-cell labeling under different experimental conditions to ensure efficient probe binding. Subsequently, LC-MS/MS mass spectrometry analysis was performed on the enriched peptide fractions, and competitive elution assays were used to identify multiple covalently bound kinase targets. The results demonstrated that the probe specifically formed covalent bonds with multiple protein kinases, with primary targets involved in cellular signaling pathways and metabolic regulation. Furthermore, biochemical experiments and computational modeling revealed that the probe’s binding to kinases depended on specific reactive amino acid residues, affecting their catalytic activity. The study concludes that LC-MS-based chemical proteomics enables efficient and precise identification of the direct targets of covalent probes. This method serves as a valuable tool for covalent drug discovery and kinase-targeted research, offering new strategies for optimizing covalent inhibitors.
Mccloud, R L. et al. Analytical Chemistry, 2021.
Figure 2. LC-MS/MS Detection of Target Proteins and Probe Modification Sites.
Deliverables
1. Comprehensive Experimental Details
2. Materials, Instruments, and Methods
3. Data Analysis, Preprocessing, and Estimation
4. Bioinformatics Analysis
5. Raw Data Files
MtoZ Biolabs, an integrated Chromatography and Mass Spectrometry (MS) Services Provider, provides advanced proteomics, metabolomics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our ultimate aim is to provide more rapid, high-throughput, and cost-effective analysis, with exceptional data quality and minimal sample consumption. Free project evaluation, welcome to learn more details!
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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