Improving the Yield of Basic Amino Acids in a Protein Sequencer

User Benefits

- Enables the use of simple pretreatment steps to improve the yield of basic amino acids from Edman degradation. - Improves yield of basic amino acids and simplifies subsequent automated software-based estimations. - Enhances yield of PTH-amino acids to improve the reliability of amino acid sequencing.

Introduction

Biopharmaceuticals are drugs developed and manufactured using biotechnological techniques, including protein and antibody drug products. Biopharmaceutical production involves multiple processes, including manufacture, purification, formulation development, and storage. Quality assurance of biopharmaceuticals requires not only quality tests on the final product but must also address the impact of raw materials and the manufacturing process. As a result, manufacturing and quality controls for biopharmaceuticals differ from those for small-molecule pharmaceuticals produced by chemical synthesis. Guidelines established for evaluating the quality of biopharmaceuticals call for the characterization of various aspects of production, including N-terminal amino acid sequencing of the protein’s primary structure. This characterization is used to compare the N-terminal amino acid sequence of the manufactured biopharmaceutical against the N-terminal amino acid sequence encoded in the gene sequence. Edman degradation is a process that sequentially removes single amino acids from the N-terminus of proteins and is used to determine amino acid sequences with high reliability. The PPSQ-50A system is a protein sequencer that automates this Edman degradation method and provides a straightforward means of determining the N-terminal amino acid sequence of proteins and peptides. The derivatives of amino acids removed by Edman degradation are called PTH- amino acids, and yields of these PTH-amino acids differ depending on the amino acid removed. Yields from basic amino acids (arginine, histidine, lysine) can be particularly low depending on the amino acid sequence and the sample, and these low yields can impact the estimation of the amino acid sequence. This article describes the development of a method that improves the yield of basic amino acids from Edman degradation.

12 de abril de 2023 GMT