Preparation method for monoclonal antibody library of tumor mucoprotein glycopeptide epitope

A monoclonal antibody, antigen-determined technology, applied in the fields of genetic engineering and immunology
CN102321923AInactive Publication Date: 2012-01-18JIANGSU UNIV +1
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
JIANGSU UNIV
Publication Date
2012-01-18
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a preparation method for a monoclonal antibody library of tumor mucoprotein glycopeptide epitope. The method comprises the following steps of: extrapolating all possibly occurring tumor glycopeptide sequences according to an amino acid sequence of mucoprotein; then transfecting tumor cells by using mucoprotein genes to obtain immunogens for immuning a mice to prepare monoclonal antibodies; synthesizing all extrapolated glycopeptides by using a chemical synthesis method and labeling with biotins respectively to obtain the biotin-labeled glycopeptides for screening the monoclonal antibodies; and finally, immuning the mice by using the immunogens to prepare hybridoma, screening the hybridoma by using the biotin-labeled glycopeptides, and sorting out the monoclonal antibody aiming at each specific glycopeptide structure. In the invention, an immune glycomic method is used for the first time to extrapolate all possibly occurring epitopes of highly glycosylated tandem repeat polypeptide segments during cancerization. The glycopeptide antigens are obtained by using the chemical synthesis method; and then the synthesized glycopeptides are used for tagging the monoclonal antibodies.
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Description

technical field

[0001] The invention belongs to the fields of genetic engineering and immunology, and in particular relates to a method for preparing a monoclonal antibody library of a tumor mucin glycopeptide epitope. Background technique

[0002] Mucin (MUCIN) is a type 1 glycoprotein expressed by epithelial cells, highly expressed on the cell surface, and highly secreted. The mucin family has nineteen members: MUC1, MUC2, MUC3A, MUC3B, MUC4, MUC5AC, MUC5B, MUC6, MUC7, MUC8, MUC12, MUC13, MUC15, MUC16, MUC17, MUC19, and MUC20. Their function is to protect the epithelial cells of the digestive tract, respiratory tract, and reproductive tract, and to regulate cell communication and signaling of epithelial cells.

[0003] The structure of mucin consists of intracellular domains, transmembrane domains, and extracellular domains. The extracellular domain can be hydrolyzed by proteases and secreted into the blood circulation. The extracellular region consists of highly glycos...

Claims

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