As the development of biotechnology, peptide and protein substances are attracting more and more attention in chemical, pharmaceutical, environmental protection fields. Recent years, as the improvement of MS technology, it has become more and more important in polypeptide amino acid sequence analysis.
In this post, we are going to introduce the sequencing analysis method of polypeptide amino acids.
Polypeptide amino acids sequence analysis is a field of proteomics. In the fields of biochemistry and cell biology, 1D or 2D sds page is usually used in isolating proteins. And now the 2D PAGE has become the most efficient method to isolate samples with at least 1000 proteins.
At the end of 80s, MALDI-TOF-MS and ESI-MS stepped into the new field of biological macromolecules. MALDI-TOF-MS can be utilized to analyze almost proteins, especially the mixture of polypeptide and proteins. And ESI-MS is the most soft ionization technique, used in various types of mass analyzers. What’s more, it can be used in online isolation and identification.
Two kinds of mass spectrometry methods
There are two methods in protein analysis. They are MS charting and MS mapping. The first one refers to component analysis, which aims to issues, body fluids or its extracts to recognize as many components of polypeptide and proteins as possible. The later one means the composition analysis. It focusses on single, purified peptides or proteins to determine its primary structure, identification of genetic differences and protein modifications, etc.
Steps of MS charting
1.Extracting polypeptides and proteins from biological tissues and segmentate extract.
2.Determine the molecular weight of each component in liquid extract.
3.Searching in peptide or protein database.
4.Analyzing and confirming again
Steps of MS mapping
1.Purifying protein samples
2.Cutting proteins with enzymatic or chemical method
3.Analyzing product by mass spectrometry
4.Using mass spectrometry database
5.Obtaining sequence information by MS/MS technology
These are some amino acid sequence analysis method in polypeptides and proteins.
In this post, we are going to introduce the sequencing analysis method of polypeptide amino acids.
Polypeptide amino acids sequence analysis is a field of proteomics. In the fields of biochemistry and cell biology, 1D or 2D sds page is usually used in isolating proteins. And now the 2D PAGE has become the most efficient method to isolate samples with at least 1000 proteins.
At the end of 80s, MALDI-TOF-MS and ESI-MS stepped into the new field of biological macromolecules. MALDI-TOF-MS can be utilized to analyze almost proteins, especially the mixture of polypeptide and proteins. And ESI-MS is the most soft ionization technique, used in various types of mass analyzers. What’s more, it can be used in online isolation and identification.
Two kinds of mass spectrometry methods
There are two methods in protein analysis. They are MS charting and MS mapping. The first one refers to component analysis, which aims to issues, body fluids or its extracts to recognize as many components of polypeptide and proteins as possible. The later one means the composition analysis. It focusses on single, purified peptides or proteins to determine its primary structure, identification of genetic differences and protein modifications, etc.
Steps of MS charting
1.Extracting polypeptides and proteins from biological tissues and segmentate extract.
2.Determine the molecular weight of each component in liquid extract.
3.Searching in peptide or protein database.
4.Analyzing and confirming again
Steps of MS mapping
1.Purifying protein samples
2.Cutting proteins with enzymatic or chemical method
3.Analyzing product by mass spectrometry
4.Using mass spectrometry database
5.Obtaining sequence information by MS/MS technology
These are some amino acid sequence analysis method in polypeptides and proteins.