Generation of recombinant antibody fragments with toxin-neutralizing potential in loxoscelism.
Loxosceles spider bites often lead to serious envenomings and no definite therapy has yet been established. In such a context, it is of interest to consider an antibody-based targeted therapy. We have previously prepared a murine monoclonal IgG (LiMab7) that binds to 32-35kDa components of Loxosceles intermedia venom and neutralizes the dermonecrotic activity of the venom. Here, we re-engineered LiMab7 into a recombinant diabody. The protein was produced in bacteria and then it was functionally characterized. It proved to be efficient at neutralizing sphingomyelinase and hemolytic activities of the crude venom despite the slightly altered binding kinetic constants and the limited stability of the dimeric configuration. This is the first report of a specific recombinant antibody for a next-generation of Loxosceles antivenoms.
Universidade Federal do Parana, Departamento de Patologia Basica, Laboratorio de Imunoquimica, Curitiba, PR, Brazil.
Sabrina
S
Karim-Silva
Universidade Federal do Parana, Departamento de Patologia Basica, Laboratorio de Imunoquimica, Curitiba, PR, Brazil.
Juliana de
Jd
Moura
University Paris-Sud, School of Pharmacy, UMS IPSIT, 5 rue J.-B. Clément, 92296 Châtenay-Malabry, France.
Magali
M
Noiray
Centro de Producao e Pesquisa de Imunobiologicos - CPPI, Piraquara, PR, Brazil.
Joao Carlos
JC
Minozzo
Université de Tours - INRA, UMR 1282, Faculté de Pharmacie, 31 Avenue Monge, 37200 Tours Cedex, France.
Nicolas
N
Aubrey
Universidade Federal do Parana, Departamento de Patologia Basica, Laboratorio de Imunoquimica, Curitiba, PR, Brazil; Muséum National d'Histoire Naturelle et Centre National de la Recherche Scientifique, UMR 7245, 57 rue Cuvier, 75231 Paris Cedex 05, France. Electronic address: lmalvarenga@ufpr.br.
Larissa M
LM
Alvarenga
University Paris-Sud, School of Pharmacy, UMS IPSIT, 5 rue J.-B. Clément, 92296 Châtenay-Malabry, France; Muséum National d'Histoire Naturelle et Centre National de la Recherche Scientifique, UMR 7245, 57 rue Cuvier, 75231 Paris Cedex 05, France. Electronic address: philippe.billiald@u-psud.fr.
Philippe
P
Billiald
D000818:Animals; D000911:Antibodies, Monoclonal / Q000627:therapeutic use*; D000997:Antivenins / Q000627:therapeutic use*; D002478:Cells, Cultured; D015870:Gene Expression; D006461:Hemolysis; D007167:Immunotherapy / Q000379:methods*; D051379:Mice; D010727:Phosphoric Diester Hydrolases / Q000276:immunology*; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000627:therapeutic use*; D057127:Single-Chain Antibodies / Q000627:therapeutic use*; D013108:Sphingomyelin Phosphodiesterase / Q000037:antagonists & inhibitors; D001098:Spider Bites / Q000276:immunology / Q000628:therapy*; D013111:Spider Venoms / Q000276:immunology*; D013112:Spiders / Q000276:immunology
D000911:Antibodies, Monoclonal; D000997:Antivenins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D013111:Spider Venoms; C055161:loxosceles venom; D010727:Phosphoric Diester Hydrolases; D013108:Sphingomyelin Phosphodiesterase
Antivenom; Diabody; Loxosceles; Neutralization; Spider; Venom