How Is the Amino Acid Sequence of a Peptide Determined?

Peptides are large biological molecules composed of amino acids. The determination of their amino acid sequence is of great significance for drug development in the field of biotechnology. This article will introduce the methods of determining the amino acid sequence of peptides and its application in drug development.

 

Edman Degradation Method

The Edman degradation method is one of the earliest methods used to determine the amino acid sequence of peptides. This method partially hydrolyzes peptides with acidic reagents, converts the N-terminal amino acid into a detectable derivative, and then separates and identifies the derivatives through techniques such as high-performance liquid chromatography to determine the type of N-terminal amino acid. Then, hydrolysis with acid is performed again, converting the second amino acid into a detectable derivative. This cycle is repeated to gradually determine the amino acid sequence of the peptide.

 

Mass Spectrometry

Mass spectrometry is a commonly used method for determining the amino acid sequence of peptides. The peptide is ionized and fragmented by a mass spectrometer to obtain the mass-to-charge ratio information of the peptide fragments. Then, through the analysis of the mass spectrometer, the amino acid sequence of the peptide can be inferred. Mass spectrometry has the advantages of high sensitivity and high resolution, and can determine the amino acid sequence of complex peptides.

 

Gene Sequencing Method

With the development of biotechnology, gene sequencing methods have also been applied to the determination of peptide amino acid sequences. The gene sequence of the peptide is determined by cloning and expressing the peptide-encoding gene and using DNA sequencing technology. Then, through transcription and translation, the amino acid sequence of the peptide is obtained. The gene sequencing method has the characteristics of high throughput and high accuracy, and is suitable for large-scale peptide amino acid sequence determination.

 

Application

The determination of the amino acid sequence of peptides has a wide range of applications in drug development. Firstly, by determining the amino acid sequence of peptides, the structure and function of peptides can be determined, providing an important basis for drug design and optimization. Secondly, the determination of the amino acid sequence of peptides can also be used to identify and control the quality of peptide components in drugs, ensuring the quality and safety of drugs. In addition, the determination of the amino acid sequence of peptides can also be used to study the biological activity and metabolic pathway of peptides, providing a theoretical basis for the clinical application of drugs.

 

The determination of the amino acid sequence of peptides is an important link in drug development in the field of biotechnology. The Edman degradation method, mass spectrometry, and gene sequencing method are commonly used methods for determining the amino acid sequence of peptides. By determining the amino acid sequence of peptides, the structure and function of peptides can be revealed, providing an important basis for drug development. The determination of the amino acid sequence of peptides has a wide range of application prospects in drug development, and will provide strong support for the discovery and development of new drugs.