A method and its application for de novo sequencing of monoclonal antibodies
CN117607307A · Shenzhen Institute for Drug Control · priority 2023
| Patent | CN117607307A ↗ |
| Status | Granted |
| Priority date | 2023-12-06 |
| Filed | 2023-12-06 |
| Published | 2024-02-27 |
| Assignee | Shenzhen Institute for Drug Control (printed Shenzhen Institute For Drug Control (shenzhen Testing Center Of Medical Devices)) |
| Also published as | CN117607307B |
Abstract
The invention provides a method for de novo sequencing of monoclonal antibodies and application thereof, wherein the method comprises the following steps: (1) Carrying out denaturation reduction and alkylation on a monoclonal antibody sample in sequence to obtain an alkylation reaction solution, dividing the alkylation reaction solution into 5 parts averagely, respectively adding trypsin, chymotrypsin, pepsin, elastase and Glu-C enzyme for enzymolysis, and mixing and reacting 5 parts of enzymolysis solution to obtain a polypeptide sample to be detected; (2) Detecting the polypeptide sample to be detected by using high performance liquid chromatography and mass spectrometry to obtain mass spectrum data of the polypeptide sample; the peptide sequences were reassembled into monoclonal antibody sequences using de novo sequencing software for analysis. The method can rapidly perform quality evaluation on the corresponding antibody, accurately detect the mutation site of the antibody, greatly simplify the quality evaluation and detection operation of the antibody reagent and shorten the quality evaluation time of the antibody.
Why it is here
Shenzhen Institute for Drug Control: de novo sequencing of monoclonal antibodies from five-protease digests.
Inventors (15)
林骏, 纪元, 刘尧, 李玉萍, 何先萍, 胡告, 邓伏雨, 杨娟, 梁永羿, 冼嘉华, 吴洪波, 张玄妮, 王振宇, 朱小林, 古慧贤
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Catalogued papers it cites (2)
- Use of PASEF for Accelerated Protein Sequence Confirmation and De Novo Sequencing with High Data Quality (2022) added by the examiner
- De novo assisted AFB1-Specific monoclonal antibody sequence assembly and comprehensive molecular characterization (2022) added by the examiner
Matched from the patent’s 6 literature citations by the rules the paper citation graph uses.
Catalogued patents it cites (3)
- CN116779037B A method for antibody sequence identification based on de novo sequencing and homology tag graph theory
- CN106645456B Sequencing method for realizing monoclonal antibody drug amino acid sequence based on mass spectrometry technology
- CN113933420A Mass spectrum-based monoclonal antibody head-on sequencing method