Amino Acid Sequence Analysis
Amino acid sequence analysis is a technique used to determine the order of amino acids in protein molecules. The structure and function of proteins are directly determined by their amino acid sequence, with amino acids linked by peptide bonds in a specific order to form protein molecules with distinct three-dimensional structures. The key advantages of amino acid sequence analysis include high sensitivity, high resolution, and high throughput. Mass spectrometry, in particular, is well-suited for analyzing complex samples, as it allows for rapid acquisition of substantial sequence information. Additionally, this technique can identify post-translational modifications (such as phosphorylation and glycosylation), which are essential for understanding protein regulation. Amino acid sequence analysis enables scientists to reveal primary structural information, predict three-dimensional protein structures, and gain deeper insights into biological functions and mechanisms. In medical research, amino acid sequence analysis plays a crucial role in the discovery of disease biomarkers and the development of protein-based drugs. For example, diseases like sickle cell anemia are caused by a single amino acid mutation in the protein sequence. This analysis can pinpoint the mutation site, uncover the molecular mechanisms of the disease, and guide personalized treatments. In biopharmaceuticals, this analysis serves as a method for quality control of protein drugs. For instance, monoclonal antibodies and recombinant protein drugs must maintain the integrity and consistency of their amino acid sequences during production. High-precision sequence analysis verifies that the protein drug sequence matches the designed sequence, ensuring drug safety and efficacy.
The fundamental principle of amino acid sequence analysis involves the stepwise degradation of the protein into individual amino acid residues using chemical or enzymatic methods. High-resolution mass spectrometry or other analytical techniques are then used to identify and sequence these amino acids. The main methods currently used in amino acid sequence analysis include Edman degradation and mass spectrometry (MS).
Edman degradation is a classic method for determining amino acid sequences. It involves reacting phenylisothiocyanate (PITC) with the N-terminal amino acid of a peptide chain, sequentially releasing and identifying individual amino acids from the N-terminus. While this method offers high accuracy, it is typically used for analyzing relatively short sequences, generally 20-30 amino acids in length.
Mass spectrometry (MS), particularly tandem mass spectrometry (MS/MS), is one of the most widely used techniques for amino acid sequence analysis. In MS, protein samples are first digested into shorter peptide fragments (typically using trypsin). These peptides are then introduced into a mass spectrometer, where they are ionized via electrospray ionization (ESI) or matrix-assisted laser desorption/ionization (MALDI). In tandem MS, the peptides are further fragmented, and the resulting ion fragment spectra are used to deduce the amino acid sequence of the peptides. Mass spectrometry not only offers high throughput and sensitivity but also enables the analysis of complex peptide mixtures.
MtoZ Biolabs is dedicated to providing efficient and accurate protein sequence analysis services to research institutions, pharmaceutical companies, and the food industry. Our technical team offers customized solutions based on various sample types and research needs, supporting clients in achieving breakthrough progress in scientific research, drug development, and industrial applications.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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