Protein Identification Service by Tandem Mass Spectrometry

Protein tandem mass spectrometry (MS/MS) is an efficient, precise, and sensitive method for qualitative and quantitative identification of proteins based on mass spectrometry technology. It is widely used in proteomics research, biopharmaceutical quality control, disease biomarker discovery, and other fields. By analyzing the peptide fragments (products of protein digestion) using mass spectrometry, this technique identifies target proteins, including unknown ones. Originating in the early 1990s, it was initially dominated by liquid chromatography coupled with mass spectrometry (LC-MS/MS). With the development of techniques such as matrix-assisted laser desorption/ionization (MALDI), protein mass spectrometry identification entered a new era. Today, MS/MS has become an important tool in proteomics research, with significant implications for deciphering protein structures and functions in complex biological samples. It aids in uncovering protein interactions, signal transduction mechanisms, and drives advancements in biological science and disease treatment.

Analysis Workflow

1. Sample Preparation

Purify, concentrate, and desalt the provided protein samples from customers using appropriate methods such as high-performance liquid chromatography (HPLC) or gel electrophoresis to remove impurities and enhance protein purity.

2. Protein Digestion

Perform multiple enzymatic digestions on purified proteins to cleave them into peptide fragments, facilitating subsequent mass spectrometry analysis.

3. Peptide Separation

Separate peptide fragments using nano-high-performance liquid chromatography (nano-HPLC) to enhance resolution and detection sensitivity effectively.

4. MS/MS Analysis

Employ high-resolution, high-sensitivity mass spectrometers (Thermo Fisher's Orbitrap Fusion Lumos, Orbitrap Exploris 240) to analyze the quality and fragment ions of separated peptide fragments through MS/MS.

5. Data Analysis

Utilize specialized data processing software such as Mascot and protein databases for peptide quality matching and fragment ion searching, ultimately achieving protein identification, including detailed information such as protein name, amino acid sequence, and matching score.

Applications

1. High-Throughput Protein Identification

MS/MS plays a critical role in proteomics research, identifying and quantifying proteins in biological samples, revealing differences in protein expression, interactions, and functions.

2. Post-translational Modification Studies

Used for identifying post-translational modifications of proteins (such as phosphorylation, acetylation, glycosylation), revealing the impact of modifications on protein function and signaling.

3. Protein Interaction Studies

By identifying components of protein complexes, it reveals the interaction relationships between proteins, providing a basis for the study of biological signaling pathways.

4. Protein Structure Analysis

MS/MS technology can analyze the primary structure (amino acid sequence) and some secondary and tertiary structure information of proteins, providing reference data for protein structure research.

5. Disease Diagnosis and Biomarker Research

MS/MS technology can be used for the identification and quantification of disease-related proteins, providing a basis for disease diagnosis and prognosis assessment.

Sample Submission Requirements

Deliverables

In the technical report, MtoZ Biolabs will provide you with a detailed report including: