Can mass spectrometry be used for protein sequencing? - Biotec Biotech

Can mass spectrometry be used for protein sequencing? - Biotec BiotechIntroduction to Testing Technology Services Details

1、 Background introduction

Sequencing can directly reveal the amino acid sequence of d à nb á izh ì, which is the basis for understanding the structure and function of d à nb á izh ì. After protein sequencing, the three-dimensional structure of d à nb á izh ì can be further predicted, and its biological functions and interaction mechanisms can be analyzed. This is of great significance for revealing the mysteries of life activities and understanding the occurrence and development of diseases. The current protein sequencing technologies mainly include: mass spectrometry de novo sequencing, Edelman degradation, and nanopore sequencing. The currently widely used method is de novo sequencing based on mass spectrometry. De novo protein sequencing is a novel sequencing technique that directly determines the amino acid sequence of d à nb á izh ì without relying on any known sequences or d à nb á izh ì database information. This technique is particularly useful for identifying novel proteins. Antibodies, as molecules in our immune system, are valuable tools in medical diagnosis and treatment. De novo sequencing has important applications in the field of antibodies.

2、 Technical process

Mass spectrometry based d à nb á izh ì sequencing uses a mass spectrometer to collect quality information of peptide amino acid sequences, namely secondary spectra. The peptide sequence is directly analyzed from the secondary spectra using de novo sequencing algorithms, and after sequence splicing, the complete d à nb á izh ì amino acid sequence is obtained. The technical process of zh ì p ǔ f ǎ d à nb á izh ì sequencing is as follows: sample preparation → d à nb á izh ì enzymatic hydrolysis → peptide separation → mass spectrometry detection → data analysis. The precautions for sequencing include: ① During sample preparation, try to avoid degradation of d à nb á izh ì; ② Select appropriate enzymes for d à nb á izh ì enzymatic hydrolysis; ③ Ensure stable performance of the mass spectrometer and improve data accuracy.

3、 Research case series

Monoclonal immunoglobulin disease (MGUS) is a plasma cell disease characterized by the presence of a major monoclonal antibody (M protein) in the serum. The author used liquid chromatography-mass spectrometry analysis to detect IgG1 type M protein in human serum. Multiple proteases were used to cleave M protein, and antibody peptides were obtained by de novo sequencing. After peptide splicing, the complete heavy and light chain sequences of M protein were finally determined. In addition, the author also characterized the glycosylation level in the CDR region of antibodies.

Mass spectrometry sequencing can help discover new drug targets and provide critical information for drug design and screening. Through sequencing technology, scientists can identify disease-related d à nb á izh ì and design drugs targeting these d à nb á izh ì to achieve targeted treatment. With the help of sequencing technology, doctors can more accurately assess patients' disease status, predict disease progression trends, and develop more personalized treatment plans. This can not only improve treatment effectiveness, but also reduce side effects and enhance patients' quality of life.

Beijing Baitai Pike Biotechnology Co., Ltd. has developed a software for antibody sequence analysis based on literature published in Nature Biotechnology and the proteomics journal JPR, combined with existing background knowledge. It has a self-developed method for calculating mass spectrometry raw data and adopts the principle of wide in and narrow out, achieving accurate identification of antibody/protein sequences without missing any effective data information. During the sequencing process, the currently high-resolution and highly sensitive Orbitrap Eclipse and Orbitrap Fusion Lumos mass spectrometers will be used to ensure data quality and accuracy of results. The protein/antibody sequencing project has a cycle of 3 to 4 weeks.

4、 Reference citation

[1]albertj,denboer,arjand,et al.Direct Mass Spectrometry-Based Detection and Antibody Sequencing of Monoclonal Gammopathy of Undetermined Significance from Patient Serum: A Case Study[J]. Journal of Proteome Research, 2023, 22(9):3022-3028.

[2]Bandeira N, Pham V, Pevzner P, et al. Automated de novo protein sequencing of monoclonal antibodies [J]. Nature Biotechnology, 2008, 26(12):1336-1338

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!

Can mass spectrometry be used for protein sequencing? - Biotec BiotechIntroduction to Testing Technology Services Details