Q-TOF Peptide Mapping Analysis Steps

Q-TOF (Quadrupole-Time of Flight) mass spectrometer is a device that combines quadrupole and time-of-flight mass spectrometry (TOF) technologies. It integrates the selective ion transmission of quadrupole mass spectrometry with the high resolution and high precision measurement capabilities of time-of-flight mass spectrometry. It is commonly used in peptide mass spectrometry analysis, particularly in protein identification and proteomics research. Q-TOF mass spectrometry is characterized by high mass accuracy and high resolution in peptide measurement.

Figure 1

The basic steps of Q-TOF peptide mapping analysis are roughly as follows:

1. Sample Preparation:

• Protein extraction: Extract proteins from biological samples.

• Protein digestion: Use enzymes (such as trypsin) to digest proteins into peptides.

2. Peptide Separation:

• HPLC: Choose suitable chromatographic columns and gradient conditions to separate mixed peptides through high-performance liquid chromatography (HPLC).

3. Mass Spectrometry Analysis:

• Peptide ionization: In the Q-TOF ion source, peptides are ionized and converted into gaseous ions.

• Precursor ion selection: The quadrupole selects specific precursor ions to enter the collision cell.

• Collision-induced dissociation: Precursor ions are energized and dissociated in the collision cell to generate fragment ions.

• Time-of-flight analysis: Measure the m/z values of fragment ions using the TOF analyzer.

4. Data Processing and Analysis:

• Peptide identification: Identify peptide sequences by matching fragment ion spectra of peptides with predicted or database spectra.

• Protein identification: Identify corresponding proteins by linking identified peptides.

• Quantitative analysis: For quantitative experiments, compare the abundance changes of peptides under different samples or conditions.

• Bioinformatics analysis: Deeply explore the biological significance of proteins and modifications.

5. Results Verification:

(1) Verify whether the identified proteins are consistent with the experimental background and purpose.

(2) Conduct further verification experiments on important proteins or peptides, such as re-conducting mass spectrometry analysis or other biochemical experiments.

Bioyork Biotech - Characterization of biologics, high-quality mass spectrometry detection services for multi-omics.

Related services:

HPLC Peptide Mapping Analysis

HPLC Quality Peptide Mapping Analysis

Peptide Coverage/Peptide Mapping Analysis

Peptide Mass Fingerprinting