3D modeling and characterization of the human CD115 monoclonal antibody H27K15 epitope and design of a chimeric CD115 target.
The humanized monoclonal antibody H27K15 specifically targets human CD115, a type III tyrosine kinase receptor involved in multiple cancers and inflammatory diseases. Binding of H27K15 to hCD115 expressing cells inhibits the functional effect of colony-stimulating factor-1 (CSF-1), in a non-competitive manner. Both homology modeling and docking programs were used here to model the human CD115 extracellular domains, the H27K15 variable region and their interaction. The resulting predicted H27K15 epitope includes mainly the D1 domain in the N-terminal extracellular region of CD115 and some residues of the D2 domain. Sequence alignment with the non-binding murine CD115, enzyme-linked immunosorbent assay, nuclear magnetic resonance spectroscopy and affinity measurements by quartz crystal microbalance revealed critical residues of this epitope that are essential for H27K15 binding. A combination of computational simulations and biochemical experiments led to the design of a chimeric CD115 carrying the human epitope of H27K15 in a murine CD115 backbone that is able to bind both H27K15 as well as the murine ligands CSF-1 and IL-34. These results provide new possibilities to minutely study the functional effects of H27K15 in a transgenic mouse that would express this chimeric molecule.
TRANSGENE S.A.; Illkirch-Graffenstade, France.
Benoît
B
Grellier
TRANSGENE S.A.; Illkirch-Graffenstade, France.
Fabrice
F
Le Pogam
NMRTEC S.A.S.; Illkirch-Graffenstaden, France.
Marc
M
Vitorino
NMRTEC S.A.S.; Illkirch-Graffenstaden, France.
Jean-Philippe
JP
Starck
TRANSGENE S.A.; Illkirch-Graffenstade, France.
Michel
M
Geist
TRANSGENE S.A.; Illkirch-Graffenstade, France.
Vanessa
V
Duong
TRANSGENE S.A.; Illkirch-Graffenstade, France.
Hélène
H
Haegel
TRANSGENE S.A.; Illkirch-Graffenstade, France.
Thierry
T
Menguy
TRANSGENE S.A.; Illkirch-Graffenstade, France; ElsaLys Biotech; Illkirch-Graffenstaden, France.
Jean-Yves
JY
Bonnefoy
TRANSGENE S.A.; Illkirch-Graffenstade, France.
Jean-Baptiste
JB
Marchand
TRANSGENE S.A.; Illkirch-Graffenstade, France.
Philippe
P
Ancian
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D019295:Computational Biology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D007378:Interleukins / Q000276:immunology*; D016173:Macrophage Colony-Stimulating Factor / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008956:Models, Chemical; D011485:Protein Binding; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D016415:Sequence Alignment
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007135:Immunoglobulin Variable Region; D007378:Interleukins; D011993:Recombinant Fusion Proteins; C552364:interleukin-34, mouse; D016173:Macrophage Colony-Stimulating Factor; D016186:Receptor, Macrophage Colony-Stimulating Factor
CD115; CSF-1; IL-34; NMR; epitope mapping; homology modeling; protein-protein docking; quartz crystal microbalance