| 摘要 |
Described is a method of selecting a protein variant having reduced immunogenicity as compared with the parent protein comprising the steps of screening a random peptide display package library with antibodies raised against any protein of interest, sequencing the amino acid sequence of the antibody binding peptides, or the DNA sequence encoding the antibody binding peptides, identifying epitope patterns of a protein by sequence alignment of the reactive peptide sequence, localisation of epitope patterns on the primary 3-dimensional structure of the parent protein, defining an epitope area including amino acids situated within 5 angstrom from the epitope amino acids, and affecting antibody binding to the epitope, changing the localised epitope patterns, or amino acids defining the epitope area of the parent protein by genetic engineering mutations of a DNA sequence encoding the parent protein without impairing functionality of the protein using the emerging epitope database for eliminating amino acid substitutions, creating new or duplicating existing epitope patterns, introducing the mutated DNA sequence into a suitable isolated human host cell, culturing the host cell and expressing the protein variant, and evaluating the immunogenicity of the protein variant using the parent protein as reference. Also described is the protein variant and use thereof, as well as to a method for producing the protein variant. |
