Fine epitope mapping of humanized anti-IgE monoclonal antibody omalizumab.
Omalizumab is a humanized anti-IgE antibody that inhibits IgE binding to its receptors on mast cells and basophils, thus blocking the IgE-mediated release of pharmacologic mediators from these cells. Previous studies have indicated that omalizumab binds to the Cepsilon3 domain of IgE, which is the binding site of IgE receptors, but the precise epitope recognized by omalizumab is unknown. In this study, we employed the phage display peptide library technology to select peptides binding to omalizumab. A striking peptide sequence motif was recovered, which is homologous to the sequence (424)HLP(426) within the Cepsilon3 domain of IgE-Fc. Our results further indicated that omalizumab specifically bound to the synthesized peptide "(421)THPHLPRALMRS(432)" containing the (424)HLP(426) motif in IgE-Fc. We therefore conclude that the (424)HLP(426) motif is the omalizumab epitope. This epitope overlaps with the high-affinity IgE receptor-binding site, thus providing insights into the structural basis for the mechanism of action of omalizumab.
Zheng
Lei
L
State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, People's Republic of China.
Li
Bohua
B
Qian
Weizhu
W
Zhao
Lei
L
Cao
Zhiguo
Z
Shi
Shu
S
Gao
Jie
J
Zhang
Dapeng
D
Hou
Sheng
S
Dai
Jianxin
J
Wang
Hao
H
Guo
Yajun
Y
D020816:Amino Acid Motifs; D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal / Q000276:immunology*; D061067:Antibodies, Monoclonal, Humanized; D018604:Epitope Mapping; D006801:Humans; D007073:Immunoglobulin E / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000069444:Omalizumab; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D007141:Immunoglobulin Fc Fragments; D019151:Peptide Library; D010455:Peptides; D000069444:Omalizumab; D007073:Immunoglobulin E