How to Accurately Detect Protein Primary Structure?

Accurately determining protein primary structure is an important task in the field of bioproducts. 

 

Amino Acid Analysis

Amino acid analysis is the basis for determining the primary structure of proteins by determining the types and relative content of various amino acids in the protein. Commonly used amino acid analysis techniques include High-Performance Liquid Chromatography (HPLC) and Capillary Electrophoresis (CE). These techniques can separate and quantify the amino acids in protein samples, thereby inferring the amino acid sequence of the protein.

 

Mass Spectrometry Sequencing

Mass spectrometry sequencing technology is an efficient and sensitive method for the analysis of protein primary structure. By digesting proteins into small peptides and then using a mass spectrometer to measure the mass and fragmentation spectrum of these peptides, the amino acid sequence of the protein can be inferred. Commonly used mass spectrometry sequencing techniques include protein resolution based on MALDI-TOF and protein sequencing based on liquid chromatography mass spectrometry.

 

DNA Sequencing and Recombination Technology

The primary structure information of proteins can be obtained at the gene level using DNA sequencing and genetic engineering technology. By cloning and expressing the genes of the target protein, the target protein can be synthesized in vitro, and its amino acid sequence can be determined by sequencing. This method is commonly used in protein engineering and biopharmaceutical fields.

 

Steps and Verification

In the process of accurately determining protein primary structure, strict experimental steps and verification are required. This includes the preparation and purification of protein samples, the verification of the accuracy of amino acid analysis, the interpretation of mass spectrometry data, and quality control. At the same time, in order to improve the reliability of the results, the accuracy of the results can be ensured through repeated experiments and cross-validation of multiple analysis methods.

 

Through key technologies and steps, such as amino acid analysis, mass spectrometry sequencing technology, DNA sequencing and recombination technology, etc., the accurate determination of protein primary structure can be achieved. This provides important information for our in-depth understanding of the structure and function of proteins.