What is the Role of Protein Sequencing?
Protein sequencing, as a key method for deciphering protein structure and function, plays an indispensable role in these processes. In 1954, Sanger successfully sequenced bovine insulin, confirming for the first time the role of amino acids in protein synthesis, thus marking the beginning of the protein sequencing field. In the early 1970s, Sanger developed DNA sequencing technology, laying the foundation for the modification of protein sequences. Proteins are critical macromolecular organic compounds in living organisms, playing vital structural and functional roles.
Protein sequencing primarily involves determining the primary structure of proteins. The most commonly used techniques today include PCR-based protein sequencing, Edman degradation sequencing, and mass spectrometry sequencing. PCR-based protein sequencing is essentially gene sequencing. This method involves sequencing RNA (or DNA) expressed in cells or tissues, which is then translated into a protein sequence according to codons. Edman degradation, a chemical method, sequentially degrades amino acids from the N-terminus of the protein, identifying each amino acid using high-performance liquid chromatography. This method is limited by protein length and can only be applied to the N-terminus of the protein, making it unsuitable when the N-terminus is blocked or when amino acids undergo chemical modifications. Mass spectrometry, a technique based on the mass-to-charge ratio, consists primarily of two steps: the mass spectrometer, which ionizes protein molecules and measures their mass-to-charge ratio, and mass spectrometry data analysis, which uses computational methods to interpret ion spectra and deduce the amino acid sequence of proteins.
Accurate and rapid analysis of complete protein sequences is crucial for studying the functions of unknown proteins. Mass spectrometry is the most commonly used sequencing method. In biomedical research, tandem mass spectrometry (MS/MS) has broad applications. It can identify and quantify protein samples, study protein modifications and interactions, and explore disease mechanisms. In drug development, tandem MS can help identify drug targets, assess drug efficacy, and investigate drug metabolism and interactions with proteins. This technology is especially important in the development of protein-based drugs, including monoclonal antibodies. As a type of protein, antibody sequencing is applied in areas such as:
1. Antibody Drug Development
Accurate primary structure information of the original drug is essential for developing generic drugs. Sequencing existing antibody products is a key step in the development of antibody-based drugs or diagnostic reagents.
2. Antibody Engineering
Obtaining antibody sequences enables recombinant expression, allowing for the production of various recombinant antibody forms (e.g., IgG, scFv, Fab, sdAb) and facilitating antibody humanization.
Although tandem MS protein sequencing plays a vital role in protein research, it still faces challenges, including the analysis of complex samples, detection of low-abundance proteins, and the complexity of data interpretation. Protein sequencing is an important tool in biotechnology, helping us understand the composition and sequence of proteins and advancing fields such as drug development, disease diagnosis, and treatment. With continuous technological advancements, protein sequencing will play an increasingly important role in biotechnology.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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