title: Affinity maturation of antiHER2 monoclonal antibody MIL5 using an epitope-specific synthetic phage library by computational design.
abstract: Increased affinities mainly equal to improved biological efficacy in many cases. By now, display methods including phage library are widely exploited to obtain higher affinity antibodies. Traditional library methods mainly focus on complementary determining region mutagenesis, in which extensive screening of variant combinations as well as large library capacity is required to find higher affinity clones. In this study, based on the modeling 3D complex structure of antigen (HER2)-antibody (MIL5) complex, the key residues of contact surface were predicted and identified to guide the synthetic phage library design. Then, epitope-specific site-directed mutagenesis phage library comprised of MIL5_scFv mutants was constructed, from which a higher affinity single chain antibody (M5scFv_ph) was screened out. Following experimental results showed that the novel antibody M5scFv_ph retained superimposed epitope to the parent antibody MIL5_scFv, and possessed similar tumor growth inhibitory activity in vivo on ovarian carcinoma xenografts.
authors.affiliation: Laboratory of Immunology, Institute of Basic Medical Sciences, P.O. Box 130(3), Taiping Road #27, Beijing 100850, China.
authors.forename: Chunxia
authors.initials: C
authors.lastname: Qiao
authors.forename: Ming
authors.initials: M
authors.lastname: Lv
authors.forename: Xinying
authors.initials: X
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Geng
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Jiyan
authors.initials: J
authors.lastname: Zhang
authors.forename: Zhou
authors.initials: Z
authors.lastname: Lin
authors.forename: Jiannan
authors.initials: J
authors.lastname: Feng
authors.forename: Beifen
authors.initials: B
authors.lastname: Shen
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D002277:Carcinoma / Q000188:drug therapy / Q000473:pathology; D045744:Cell Line, Tumor; D003198:Computer Simulation; D015195:Drug Design; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008958:Models, Molecular; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000473:pathology; D019151:Peptide Library; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D047368:Tumor Burden / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D000939:Epitopes; D019151:Peptide Library; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human