Design and evaluation of a diabody to improve protection against a potent scorpion neurotoxin.
Diabodies are recombinant, dimeric, antibody-based molecules composed of two non-covalently associated single-chain antibody fragments that bind to an antigen in a divalent manner. In an attempt to develop more effective therapeutic molecules against scorpion venoms, we designed a diabody derived from monoclonal antibody 9C2, which neutralizes the toxicity of scorpion neurotoxin AahI in mammals. The recombinant diabody produced in the periplasm of Escherichia coli was purified to homogeneity in a single step by protein L-agarose affinity chromatography. It was functional, and possessed a high binding affinity to AahI (8 x 10(-11) M). The bivalence of the diabody was confirmed by size-exclusion chromatography, isoelectrofocussing and electron microscopic observations. Finally, the diabody showed high thermal stability in serum and demonstrated protective activity when injected intraperitoneally in mice experimentally envenomed with toxin AahI. In conclusion, the diabody format gives the 9C2 molecule advantageous properties that are particularly important for potential clinical applications in the treatment of envenomations.
Muséum national d'Histoire naturelle, USM 0505-Lerai, 57 rue Cuvier, 75231 Paris cedex 05, France.
N
N
Aubrey
C
C
Devaux
P Y
PY
Sizaret
H
H
Rochat
M
M
Goyffon
P
P
Billiald
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D002846:Chromatography, Affinity; D015195:Drug Design; D013058:Mass Spectrometry; D051379:Mice; D009498:Neurotoxins / Q000276:immunology* / Q000378:metabolism; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000276:immunology*; D012604:Scorpion Venoms / Q000276:immunology* / Q000378:metabolism; D012605:Scorpions / Q000378:metabolism
D000906:Antibodies; D009498:Neurotoxins; D011994:Recombinant Proteins; D012604:Scorpion Venoms; C065314:scorpion toxin I, Androctonus