Nucleic acid sequencing is now very mature, and the sequencing accuracy of most sequencers exceeds 99%. However, full-length protein sequencing (amino acid sequence) has always been a challenge, and its sequence coverage, accuracy, and cost are difficult to meet the needs of applications. In particular, the development and process establishment of biopharmaceutical preparations such as recombinant proteins or antibodies require confirmation of the full length of the protein.
In 1967, Edman and Beggs invented the Edman degradation method for protein sequencing, but it can only measure 25-50 amino acids at the N-terminal of the peptide chain, and the cost is very high. With the development of protein mass spectrometry, mass spectrometry has been increasingly used for de novo sequencing of proteins. Since the entire protein is difficult to analyze in a mass spectrometer, it needs to be degraded into short peptide fragments with enzymes in advance, and then analyzed by a mass spectrometer to identify the amino acid sequence of each peptide fragment from the spectrum, and then then splice the sequences of the multiple peptide segments into a complete protein sequence.
Applications
- Full amino acid sequence analysis of proteins, peptides, antibodies, vaccines and other biological products
- Analysis of post-translational and chemical modifications of proteins
- Quality analysis of in vitro synthesized proteins and recombinant protein expression
Our Full Length protein/Antibody Sequencing Platform
Using the existing high-resolution mass spectrometry technology platform, Creative Proteomics has developed a mass spectrometry-based protein full-sequence technology to achieve 100% coverage of the target protein sequence. Our protein/antibody full-length sequencing is based on liquid-phase mass spectrometry for full-length De Novo sequencing analysis, providing ultra-high precision protein full-length sequencing solutions with the help of the high-resolution Orbitrap platform.
What are our services used for?
This analysis service can be used to confirm whether the recombinant protein is fully expressed, and to detect whether the recombinant protein is broken during the expression process. This technology also allows fast and accurate analysis of primary sequence information for different isoforms of antibodies (e.g. IgG and IgM) and different types of antibodies (fluorescent coupled antibodies, immobilized antibodies, antibodies of different species).
What you need to do
We provide a full suite of full-length protein sequencing analysis testing services, from experiments to data interpretation to final reports. You only need to provide a protein sample to obtain accurate information about the full-length protein sequence of the sample.
Sample requirements
1. Both in-gel and solution samples can be used for the sequencing
2. Volume of target protein: 10-20ug
3. Purity >95%
Workflow of protein antibody full-length sequencing service
Delivery
1. Experimental procedure
2. Relevant mass spectrometry parameters
3. MS/MS spectra
4. Raw data
5. Full protein sequence
6. Antibody detailed result report:
Heavy chain + light chain.
1) Exact molecular weight (deglycosylated/glycosylated)
2) Amino acid sequence (100% sequence coverage)
3) The number of peptide spectrum matches
4) Peptide list of variable region
5) Leucine/isoleucine difference
