Overview of Protein De Novo Sequencing: Break Through Traditional Analysis Limitations
In proteomics and biopharmaceutical research, determining the amino acid sequence (primary structure) of proteins is essential for elucidating their functions and biological significance. However, conventional protein sequencing methods primarily rely on database searches, making them inadequate for characterizing unknown proteins, novel variants, or unannotated species. As research advances, addressing this limitation has become increasingly critical. Protein de novo sequencing, a technique that deduces protein sequences directly from mass spectrometry data without the need for reference databases, provides a novel approach to protein structural analysis.
What Is Protein De Novo Sequencing?
De novo protein sequencing is a mass spectrometry (MS/MS)-based method that determines amino acid sequences directly from peptide fragmentation spectra without reliance on existing sequence databases. Unlike traditional protein identification, which depends on previously cataloged sequences in protein databases, de novo sequencing enables the characterization of newly discovered proteins, mutated variants, and complex post-translationally modified proteins. This capability makes it particularly advantageous for studying non-model organisms, novel antibodies, and other systems lacking reference sequences.
Why Is Protein De Novo Sequencing Important?
1. Characterizing Unknown Proteins Not Represented in Databases
A vast number of proteins found in natural environments, disease-related samples, and drug discovery remain unrecorded in existing databases. De novo sequencing enables the identification and characterization of such "unknown proteins" without requiring prior sequence knowledge, making it particularly valuable for research on non-model organisms, natural bioactive compounds, and extremophilic microorganisms.
2. Determining the Authentic Sequence of Antibodies and Recombinant Proteins
For antibody therapeutics and recombinant proteins, de novo sequencing allows direct determination of the expressed product's amino acid sequence. This capability is crucial for detecting mutations, post-translational modifications, or discrepancies in expression, serving as an essential tool for quality control and consistency assessment in biopharmaceutical production.
3. Identifying Mutations and Novel Post-Translational Modifications (PTMs)
De novo sequencing facilitates the detection of previously unrecognized mutation sites and rare or complex combinatorial modifications, such as glycosylation and phosphorylation, without requiring predefined modification assumptions. This approach is instrumental in exploring disease-related mutations, drug-target interactions, and novel regulatory mechanisms in cellular biology.
How Does Protein De Novo Sequencing Differ from Traditional Database-Dependent Sequencing?
Figure 1
Applications of Protein De Novo Sequencing
Figure 2
The emergence of protein de novo sequencing represents a paradigm shift in proteomics, transitioning from the validation of known proteins to the exploration of unknown ones. By surpassing the constraints of traditional analytical frameworks, this technology not only expands the frontiers of fundamental research but also holds transformative potential in diverse fields, including disease diagnosis, drug development, and synthetic biology. Looking ahead, the integration of high-resolution mass spectrometry, artificial intelligence-driven algorithms, and more efficient sample preparation techniques will further enhance the accuracy and throughput of de novo protein sequencing. MtoZ Biolabs offers precise and rapid de novo protein sequencing services, empowering researchers with a comprehensive understanding of protein structures and functions.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
Related Services
How to order?
