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De Novo Peptide Sequencing

Principle:

De Novo Peptide Sequencing is a tech to analyze proteins or peptides not present in currently available databases.

We use tandem mass spectrometry (MALDI TOF-TOF, or electrospray MS/MS) to determine the primary sequence structure. The expert operators will then use the PEAKS Software to interpret the MS/MS spectra and derive the customer sequence.

Sample preparation:

The sample preparation for De Novo Peptide Sequencing is the same as MALDI-TOF/TOF

Application:

1. De Novo Peptide Sequencing is used to identify proteins, often on samples which failed to give a conclusive identification by peptide mass fingerprinting or for samples from species which do not have any genome sequence.

2. De Novo Peptide Sequencing is an effective tool for identification of post-translational modifications (PTM) and sequence similarity.

Service includes:

This service includes,

1) Excision of bands
2) Tryptic digestion
3) Acquisition of the peptide map of a digested protein, as well as MS/MS spectrums of 5 peptides of the most intense in the map as is the case with simple sample identification 4) An in depth interpretation of the MS/MS spectrums, so that the user receives the maximum sequencing information from the amino acids of these 5 peptides. This is especially useful in cases where the genome of the organism is not complete in the database

Creative Proteomics has developed a new technology to determine the complete amino acid sequence of monoclonal antibodies. Due to its large size and its vast diversity in antibody sequences, it used to be an extremely difficult task to carry out the de novo sequencing of such a large protein. Recently, with the breakthrough we achieved, we are capable to determine the complete sequence of a mAb with as low as 100 ug sample of moderate purity (>85%).

This new technology will have a significant impact on the discovery of novel therapeutic antibodies, as well as on the development of generic or biosimilar antibody drugs. Instead of going through a long and costly process to generate antibody from scratch, we can rapidly screen an antibody library or commercially available mAb antibodies to identify a drug candidate in an assay. This technology also provides the most conclusive way to confirm the primary amino acid sequence of therapeutic antibody product, since the conventional Peptide Mapping may fail to detect some mutated sequence.

Please contact us if interested in our mAb De novo sequencing service.