What are the reagents used in Edman degradation sequencing and its basic principles?

Edman degradation is a classic method for analyzing the N-terminal sequence of proteins. It utilizes a series of chemical reactions to cleave and identify one amino acid at a time from the peptide chain, repeating the process to determine the sequence of N-terminal amino acids.

The main reagent used in the Edman degradation method is phenyl isothiocyanate (PITC). PITC can react with the N-terminal amino acid residue of protein polypeptides to form a phenylthiocarbamoyl derivative (PTC-peptide). Under trifluoroacetic acid treatment, the peptide bond of the N-terminal amino acid is selectively cleaved, releasing a thiazolinone aniline (ATZ) derivative containing the terminal amino acid. The ATZ derivative is unstable and can be converted into a stable phenylthiohydantoin (PTH) derivative, i.e., PTH amino acid, under acidic conditions. At this point, the first amino acid at the N-terminus of the protein is obtained. The protein polypeptide, now missing one amino acid, is recovered, and the above reaction is repeated. The cleaved amino acids are identified, and the process is cycled until the N-terminal amino acid sequence determination is complete.

Biotech Pack Company uses Shimadzu's Edman sequencing system to provide a wide range of researchers and research clients withN-terminal sequencing based on Edmanservice technology packages, which can accurately determine the N-terminal amino acid sequence of purified protein products, antibodies, and protein vaccines. Additionally, Biotech Pack Biotechnology has established a platform for N-terminal sequencing using advanced LC-MS/MS technology, capable of determining blocked and modified protein termini, complementing the Edman degradation method to ensure the smooth progress of N-terminal sequencing services.

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