As a manufacturer, you want to carry out de novo sequencing of your proteins
For the biopharmaceutical industry, understanding the structure and properties of proteins is essential for innovation, quality assurance and meeting regulatory requirements. However, when proteins are unknown or little studied, conventional analysis methods based on databases are not enough. De novo sequencing is distinguished by its ability to determine the complete sequence of a protein without recourse to a reference sequence.
What is de novo sequencing?
De novo sequencing is a technique that reconstructs the amino acid sequence of a protein by directly analysing peptide fragments obtained by enzymatic cleavage. Unlike traditional methods that rely on existing databases, this approach is independent and allows work on unknown proteins.
This ability to explore the unknown makes de novo sequencing a particularly suitable protein analysis for identifying proteins from complex matrices, biomarkers or therapeutic products in development.
The steps involved in de novo sequencing
De novo sequencing involves several essential phases:
Each step is optimised to provide accurate and reliable results, even for complex or unknown proteins.
Why carry out de novo sequencing in the laboratory?
De novo sequencing is used by the FILAB laboratory to identify and characterise unknown or unrecorded proteins. This method makes it possible to:
Characterize unknown proteins: identify unreferenced sequences that are essential for the discovery of new molecules.
Optimising biomedicines: understanding variations in therapeutic proteins and improving their formulation.
Analyse post-translational modifications: detect structural modifications that are important for the function or efficacy of proteins.
Exploring biomarkers: identifying specific proteins in biological samples for diagnostic or research applications.
The FILAB laboratory provides tailor-made analysis for de novo sequencing and characterization of your proteins
Expertise in biopharmaceutical matrices
Our expertise covers a wide range of biopharmaceutical matrices, including :
- Formulated solutions and sera: for the sequencing of monoclonal antibodies and therapeutic proteins.
- Cell culture media: analysis of recombinant proteins from CHO cells or other expression systems.
Technical resources for de novo sequencing
Our expertise in mass spectrometry for de novo sequencing
The FILAB laboratory uses state-of-the-art mass spectrometry (MS/MS) technologies to analyse and reconstruct the peptide sequence of proteins. This approach is based on several key steps:
Provides a precise spectrum of peptides to analyse the structure of proteins and detect any anomalies.
Ensures reliable reconstruction of the peptide sequence, guaranteeing the integrity and conformity of the biomolecules.
Unlike peptide mass fingerprinting, which compares data with existing databases, de novo sequencing can be used to characterise unknown proteins, check the purity of bioproducts and identify potential contaminants.
Our analysis offers optimum precision for the identification of new, modified or complex proteins, with high sensitivity.
Our services
De novo sequencing allows unknown proteins to be compared with other sequences to detect similarities, mutations or modifications. This approach is useful for :
- Aligning sequences: identifying similarities between proteins or with nucleotide sequences.
- Studying the evolution of proteins: understanding their origin and functional role.
- Checking structural conformity: comparing proteins produced with those expected during biopharmaceutical development.
FILAB offers de novo sequencing analyses for monoclonal antibodies, making it possible to:
- Verify sequence integrity and conformity.
- Identify post-translational modifications, such as glycosylation.
- Guarantee the quality of products under development or in production.
Our services include the sequencing of recombinant proteins expressed in systems such as CHO cells. We help to :
- Validate sequences obtained after recombinant expression.
- Detect variations or contaminants.
- Optimise purification and formulation steps.
FILAB carries out the sequencing of specific proteins in biological samples in order to:
- Identify biomarkers linked to pathologies.
- Compare proteins in healthy and pathological matrices.
- Facilitate the development of innovative tests and diagnostics.
Our protein analysis
Protein analysis
Applications of de novo sequencing in the biopharmaceutical industry
De novo sequencing has key applications in a variety of industrial and scientific fields:
- Identification and characterization of therapeutic proteins
- Study of monoclonal antibodies
- Verification of the primary structure of recombinant proteins
- Discovery of new proteins: this enables the identification of unknown proteins, often present in complex matrices, and thus contributes to fundamental research or biopharmaceutical innovation.
- Detection of exogenous proteins or impurities in bioprocesses
- Identification of contaminants in production batches
- Confirmation of the primary structure of proteins
- Checks the integrity of biotherapies
- Analysis of post-translational modifications: this method identifies structural variations, such as phosphorylation or glycosylation, that are not recorded in databases.
- Studies on complex proteins: ideal for analysing protein mixtures from biological or environmental matrices, where conventional sequences are insufficient.
These applications demonstrate the importance of this analysis in the discovery of biomarkers or the development of biomedicines.
Advantages of de novo sequencing
De novo sequencing has a number of major advantages that set it apart from conventional approaches:
- Database independence: ideal for exploring novel or little-studied proteins.
- Precise identification of variants: detection of unexpected or undocumented structural modifications.
- Adaptability to complex matrices: effective analysis in complex or poorly purified samples.
These advantages make it an essential method for innovation projects in biopharmaceutical and biomedical research.
FAQ
De novo sequencing is used to check the integrity and sequence of proteins present in serums or formulated solutions, such as monoclonal antibodies. This ensures compliance with therapeutic specifications and detects any structural modifications or impurities.
For recombinant proteins, de novo sequencing precisely identifies the sequence obtained after expression in cell culture media. It can also detect post-translational variations and check for the absence of contaminants, thereby optimising the purification and production processes.
De novo protein sequencing is a key tool for biopharmaceuticals, enabling the identification and characterization of therapeutic proteins without the need for an existing database. This approach is essential for validating the structural integrity of biomedicines, optimising the development of new molecules and ensuring that products comply with current regulations (EMA, FDA).
The structural characterization of proteins without reference is based on de novo sequencing, which enables the primary structure of an unknown or modified protein to be analysed. This method is particularly useful for detecting variants, identifying protein impurities and controlling the purity of bioproducts, especially in the bioproduction and formulation of biomedicines.
Deciphering the primary structure of biomolecules is a key step in de novo protein sequencing, making it possible to determine the exact sequence of peptides and highlight any post-translational modifications. This analysis is crucial for optimising the stability of therapeutic proteins, ensuring the reproducibility of production batches and guaranteeing the efficacy of biopharmaceutical treatments.
Tandem mass spectrometry (MS/MS) is essential for identifying unknown proteins, monitoring the purity of biomedicines and detecting contaminants. By fragmenting peptides, it enables precise sequencing without a database, ensuring the conformity and traceability of biopharmaceutical products. Entrusting this analysis to a laboratory guarantees reliability and compliance with regulatory standards.
De novo sequencing is essential for characterizing unrecorded proteins and analysing variants, unlike peptide fingerprinting, which is limited to existing databases. For manufacturers, this approach enables more advanced quality control, reducing risks and ensuring the conformity of biomedicines. An expert laboratory offers precision, state-of-the-art equipment and regulatory support.
