Introduction
Antibody sequence information is crucial to understanding the structural basis for antigen binding and enables the use of antibodies as therapeutics and research tools. Sequencing of monoclonal antibodies contributes to the development of biopharmaceuticals and diagnostic kits. In particular, primary antibody sequence analysis is the basis for antibody development and the development of antibody drugs for humanization. However, due to the variation and modification of antibody products, the sequence information of most antibodies is often not included in the available databases. De novo sequencing technology uses advanced data processing algorithm for sequence analysis, has a superior advantage versus database dependent traditional methods. Antibody sequencing can be accurately analyzed using the de novo sequencing method, with no need for database searching.
Fig. 1. De novo protein sequencing of monoclonal antibodies. (Peng W W, et al., 2021).
Our Services
Based on advanced mass spectrometry platform, and combined with rich experience in bioinformatics analysis, Creative Proteomics has established a new generation of antibody de novo sequencing platform to achieve accurate analysis of the primary antibody structure. The method uses a panel of multiple complementary proteases to generate suitable peptides for de novo sequencing by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in a bottom-up fashion. The method achieves full sequence coverage of the monoclonal antibody, with an accuracy of 99% in the variable regions. Our customers have widely used our sequencing methods to achieve desirable results in the field of antibody drug discovery.
Advantages of Our Services
- Full Coverage of the Sequence
To achieve 100% antibody sequencing, Creative Proteomics uses a panel of multiple complementary proteases that will achieve 100% full sequence splicing of the light and heavy chains of the antibody molecule.
- Advanced Data Processing Algorithms
Creative Proteomics combined bioinformatics and developed its own mass spectrometry raw data calculation method based on the existing professional software PEAKS Mass Spectrometry Software for antibody sequence analysis. At the same time, the accurate identification of the antibody sequence was achieved.
- Detection of Sample Diversity
The technology platform we have established is not affected by small groups (such as FITC, Biotin), although larger protein groups will make sequencing difficult, but if a higher concentration of protein is provided, it can still be successfully sequenced.
- Sequence Prediction and Rigorous Sequence Validation
We will first predict the sequence of the sequencing results, indicating the CDR region of the antibody; at the same time, we will also establish a new database based on the determined sequence, and use the Sequence Confirmation mode to collect mass spectrometry data again and analysis them.
Please feel free to contact us If you would like to discuss the best way to determine the sequencing of your antibody. At Creative Proteomics, we provide professional antibody de novo sequencing services for global customers. Our scientists can also design the best solutions according to your specific requirements, and describe them in detailed project reports, we look forward to with your cooperation.
References
- Meyer L, Lopez T, Espinosa R, et al. (2019) A simplified workflow for monoclonal antibody sequencing. PLoS One. 14(6):e0218717.
