What are the two mass spectrometry methods for protein identification? - Baitai Pike

What are the two mass spectrometry methods for protein identification? - Baitai PikeIntroduction to Biotechnology Testing Technology Services Details

Identification of d à nb á izh ì is a step in d à nb á izh ì omics, and mass spectrometry (MS) is one of the core technologies for d à nb á izh ì identification. Mass spectrometry technology, by analyzing the mass to charge ratio of d à nb á izh ì molecules, can provide detailed information about d à nb á izh ì and is widely used in the identification of d à nb á izh ì. Here are two commonly used mass spectrometry methods.

1、 Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS)

1. Principle: This method first mixes the d à nb á izh ì sample with a matrix, and then forms gaseous ions between the sample and the matrix through laser irradiation. These ions then fly in an electric field, and their flight time varies depending on their mass to charge ratio (m/z), allowing for the identification of d à nb á izh ì.

2. Characteristics: MALDI-TOF MS is suitable for the analysis of larger molecular weight d à nb á izh ì, with simple operation and fast analysis speed, and is a commonly used technique in d à nb á izh ì identification.

2、 Electrospray ionization mass spectrometry (ESI-MS)

1. Principle: Electrospray ionization mass spectrometry is to form charged droplets of solution samples in a high electric field. With the evaporation of solvent, these droplets become smaller and smaller. Zu ì eventually leads to the ionization of sample molecules and is introduced into the mass spectrometer for quality analysis, so as to achieve the identification of d à nb á izh ì.

2. Characteristics: ESI-MS is suitable for analyzing polar compounds and biomolecules, especially for sequence analysis of d à nb á izh ì and peptides. It can be combined with liquid chromatography (LC) and becomes an important tool for d à nb á izh ì identification.

3、 Advantages and disadvantages of MALDI-TOF MS and ESI-MS

Advantages of MALDI-TOF MS

(1) Easy to operate: Sample preparation is relatively simple, with low requirements for samples, suitable for high-throughput analysis in d à nb á izh ì identification.

(2) Fast analysis speed: able to quickly identify d à nb á izh ì, suitable for large-scale screening.

(3) High sensitivity: It has high detection sensitivity for larger molecular weight d à nb á izh ì, especially in the identification of d à nb á izh ì.

(4) Wide applicability: It can analyze various biomolecules, such as d à nb á izh ì, peptides, nucleic acids, etc.

2. Disadvantages of MALDI-TOF MS

(1) Lower resolution: Compared to ESI-MS, MALDI-TOF MS has lower resolution and limited ability to distinguish molecules with similar mass.

(2) Limited quality range: not suitable for analyzing extremely small or large molecules.

(3) Matrix effect: The choice of matrix has a significant impact on the results, and it is necessary to optimize the ratio of matrix to sample.

3. Advantages of ESI-MS

(1) High sensitivity: capable of detecting samples at extremely low concentrations, suitable for identification of j ī ngqu è d à nb á izh ì.

(2) High resolution: capable of distinguishing molecules with very similar masses, suitable for detailed mass analysis.

(3) Wide applicability: It can be combined with separation techniques such as liquid chromatography (LC) to enhance the complexity processing capability of d à nb á izh ì identification.

(4) Sequence analysis can be performed: By using tandem mass spectrometry (MS/MS) technology, the amino acid sequence of d à nb á izh ì can be analyzed.

4. Disadvantages of ESI-MS

(1) Complex operation: Sample preparation and instrument operation are relatively complex, requiring a high level of technical expertise.

(2) Longer analysis time: Compared to MALDI-TOF MS, the analysis time is longer and not suitable for rapid high-throughput analysis.

(3) The requirements for the sample are high: the sample needs to have good purity and stability.

The combination of MALDI-TOF MS and ESI-MS can provide more comprehensive identification information for d à nb á izh ì, including its molecular weight, amino acid sequence, and post-translational modifications, thus providing a deeper understanding of the structure and function of d à nb á izh ì.

Baitai Paike Biotechnology Characteristic Project

1、 Protein sequencing

Biotec Biotech uses Thermo's newly launched Obitrap Fusion Lumos mass spectrometer and Shimadzu's Edman degradation sequencing system to analyze the d à nb á izh ì sequence, providing mass spectrometry based protein sequencing analysis services, including amino acid composition analysis of d à nb á izh ì, Ndu ā nc è x ù, Cdu ā nc è x ù, and full sequence analysis, as well as d à nb á izh ì N-terminal sequence analysis services based on Edman degradation. For unknown theoretical sequences, we provide de novo sequencing services based on de novo sequencing to analyze protein sequences.

※ Service advantages:

1. Adopting the world's advanced mass spectrometer Obitrap Fusion Lumos;

2. It can achieve 100% coverage of the measured target protein sequence;

3. It can determine up to 70 amino acid sequences at the N-terminus of proteins;

4. Can measure various forms of samples: protein solution, PVDF protein bands;

5. Low sample usage: Protein samples only require 5-10ug to complete detection;

6. Sequencing is not affected by N-terminal modifications such as blocking, PEC, and glycosylation.

2、 D à nb á izh ì omics

Biotec Biotech uses Thermo Fisher's Orbitrap Fusion Lumos mass spectrometry platform combined with Nano LC to provide a variety of d à nb á izh ì omics analysis services, including quantitative d à nb á izh ì omics, targeted d à nb á izh ì omics, proteomics, post-translational modified proteomics, and more. In addition, Baitai Paike Biotechnology has launched a 4DD à nb á izh ì omics service based on timsTOF Pro, which helps with research in micro sample proteomics, large sample population medicine, and high-throughput modification genomics.

※ Service advantages:

1 . High throughput quantitative protein analysis: Large scale experimental analysis with multiple control groups, discovering new biomarkers;

2. Multiple in vitro and in vivo labeling methods are suitable for analyzing various samples such as tissues, cells, and blood;

3. High sensitivity of mass spectrometry analysis and high repeatability of experimental results;

4. Can detect low abundance proteins with a wide linear range;

5. Bioinformatics analysis is more systematic and accurate.

3、 Single cell mass spectrometry flow cytometry analysis

Baitai Paike Biotechnology uses the Fluidigm mass spectrometry flow cytometry system for single-cell mass spectrometry analysis. Metal element markers (usually metal element labeled specific antibodies) are used to label molecules on the surface and inside of cells. Then, individual cells are separated using flow cytometry principles, and the atomic mass spectrum of individual cells is analyzed using inductively coupled plasma mass spectrometry (ICP-MS). The atomic mass spectrum data is then converted into signal molecule expression levels on the cell surface and inside.

※ Service advantages:

1. Advanced technology, filling technological gaps

The use of metal labeled antibody technology avoids the problem of few and easily interacting channels in traditional flow cytometry fluorescence. At the single-cell level, multiple indicators can be characterized simultaneously, and Baitai Paike Biotechnology can detect 51 target proteins simultaneously.

2. Large analysis quantity and low cost

Single cell RNAseq is limited by factors such as cost, and the total number of analyzed cells for all samples is generally around 2x10 ^ 4. However, flow cytometry technology can analyze at least 10 ^ 5 cells in one sample, achieving an order of magnitude increase, and the cost is not higher than that of single-cell RNAseq.

3. Great application prospects

① The results of flow cytometry can provide changes in cell subpopulations, which have great research prospects in clinical diagnosis, disease mechanism research, and other fields;

② Combining metal tag technology with other technologies will lead to new application directions. In addition to conventional proteins, mass spectrometry flow cytometry technology can also be used for post-translational modification of proteins;

③ Can detect cell survival rate, cell size, mRNA transcript expression level, DNA synthesis rate, and protease activity.

4、 Immunopeptide omics analysis and novel antigen discovery based on high-precision mass spectrometry

The one-stop solution for high-precision mass spectrometry based immunopeptidomic analysis and new antigen discovery from Biotec Biotech includes our proprietary, highly sensitive immunopeptide enrichment and identification scheme. We can assist you in identifying and recognizing over 10000 type I peptides and over 10000 type II peptides. Through our optimized high-throughput immunopeptidomic analysis platform, immunopeptidomic analysis can be performed with reproducible identification and quantification from small sample materials. This service can be applied to large-scale research, aiming to assist researchers in finding solutions for cancer, immune diseases, and infectious diseases, and to explore unknown targets in depth.

5、 Characterization of Biopharmaceuticals

Baitai Parker is based on high-resolution mass spectrometry technology, MALDI TOF， High performance chromatography separation technology provides a range of biological drug analysis solutions, from amino acid composition and primary structure analysis of biological products such as d à nb á izh ì, peptides, antibodies, vaccines, to product variability and purity analysis. Intended to provide high-quality biopharmaceutical analysis services and assist biopharmaceutical manufacturers in improving the quality of biopharmaceuticals.

Baitai Paike Biotechnology's Seven Major Testing Platforms

Baitai Paike Biotechnology - High quality service provider for characterization of biological products and multi omics of biological mass spectrometry

Beijing Baitai Paike Biotechnology Co., Ltd. is committed to providing quality control testing and project validation services for the biological/pharmaceutical and medical device industries. The company's laboratory follows regulations and guidelines such as NMPA, ICH, FDA, and EMA, and has obtained CNAS/ISO9001 dual quality system certification. It has established a complete quality system, including data cold and hot/remote backup, regular equipment measurement/periodic verification, software audit tracking, and provides customers with integrated solutions and technical services to support new drug research and development, drug application registration, and production release.

1. The company adopts the ISO9001 quality control system and provides CRO detection and analysis services based on mass spectrometry;

2. Recognized by the National CNAS Laboratory, we provide customers with drug quality research services that comply with global pharmaceutical regulations;

3. The business scope covers omics, proteomics, metabolomics, biopharmaceutical characterization, single-cell analysis, single-cell mass spectrometry flow cytometry, bioinformatics cloud analysis, and multi omics integrated analysis of biological mass spectrometry;

4. Seven major quality control testing platforms to meet your one-stop service needs;

5. Providing services to over 3000 enterprises and 10000+customers;

6. Committed to providing you with high-quality biological mass spectrometry analysis services!

What are the two mass spectrometry methods for protein identification? - Baitai PikeIntroduction to Biotechnology Testing Technology Services Details