tailieunhanh - Báo cáo khoa học: Diversity and junction residues as hotspots of binding energy in an antibody neutralizing the dengue virus

Dengue is a re-emerging viral disease, affecting approx. 100 million individ-uals annually. The monoclonal antibody mAb4E11 neutralizes the four serotypes of the dengue virus, but not other flaviviruses. Its epitope is included within the highly immunogenic domain 3 of the envelope glyco-protein E. | iFEBS Journal Diversity and junction residues as hotspots of binding energy in an antibody neutralizing the dengue virus I 1IIOQ R I I o 11 o1 I o I I I D tri R o I Izil 111 Pal rĩ C ls Pn ilĩjnrl1 I Irtolzi 1 I II I icrw a1 Hugues Dedouelle I Lauielll Delkadi I Patiiuk LliyiallU J. Inaki Guijaiio Olesia LloUVa Agathe Urvoas1 Muriel Delepierre2 and Philippe Thullier3 1 Unit of Molecular Prevention and Therapy of Human Diseases CNRS-FRE 2849 Institut Pasteur Paris France 2 Unite de RMN des Biomolecules CNRS-URA 2185 Institut Pasteur Paris France 3 Departement de Biologie des Agents Transmissibles Centre de Recherche du Service de Sante des Armees La Tronche France Keywords antibody complementary determining region dengue virus gene rearrangement molecular recognition Correspondence H. Bedouelle Unit of Molecular Prevention and Therapy of Human Diseases CNRS-FRE 2849 Institut Pasteur 28 rue Docteur Roux 75724 Paris Cedex 15 France Fax 33 1 40 61 35 33 Tel. 33 1 45 68 83 79 E-mail hbedouel@ Present address Plate-forme de Biophysique des Macromolecules et de leurs Interactions Institut Pasteur Paris France Received 17 August 2005 revised 6 October 2005 accepted 31 October 2005 doi Dengue is a re-emerging viral disease affecting approx. 100 million individuals annually. The monoclonal antibody mAb4E11 neutralizes the four serotypes of the dengue virus but not other flaviviruses. Its epitope is included within the highly immunogenic domain 3 of the envelope glycoprotein E. To understand the favorable properties of recognition between mAb4E11 and the virus we recreated the genetic events that led to mAb4E11 during an immune response and performed an alanine scanning mutagenesis of its third hypervariable loops H-CDR3 and L-CDR3 . The affinities between 16 mutant Fab fragments and the viral antigen serotype 1 were measured by a competition ELISA in solution and their kinetics of interaction by surface plasmon resonance. The .