A novel human recombinant antibody fragment capable of neutralizing Mexican scorpion toxins.
Using phage display and directed evolution, our group has progressed in the construction of a second family of human single chain variable fragments (scFv) which bind to scorpion toxins dangerous to mammals. It was observed that scFv C1 only bound initially to toxin Cn2, which constitutes 6.8% of whole venom from the scorpion Centruroides noxius Hoffman. Only a few amino acid changes were necessary to extend its recognition to other similar toxins and without affecting the recognition for its primary antigen (Cn2 toxin). One variant of scFv C1 (scFv 202F) was selected after two cycles of directed evolution against Cll1 toxin, the second major toxic component from the venom of the Mexican scorpion Centruroides limpidus limpidus Karsh (0.5% of the whole venom). scFv 202F is also capable of recognizing Cn2 toxin. Despite not having the highest affinity for toxins Cll1 (KD = 25.1 × 10(-9) M) or Cn2 (KD = 8.1 × 10(-9) M), this antibody fragment neutralized one LD50 of each one of these toxins. Additionally, scFv 202F moderately recognized Cll2 toxin which constitutes 1.5% of the venom from C. limpidus. Based on our previous experience, we consider that these results are promising; consequently, we continue working on generating new optimized variants from scFv C1 that could be part of a recombinant scorpion anti-venom from human origin, that might reach the market in the near future.
Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, UNAM, Apartado Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
Lidia
L
Riaño-Umbarila
Timoteo
T
Olamendi-Portugal
Citlalli
C
Morelos-Juárez
Georgina B
GB
Gurrola
Lourival D
LD
Possani
Baltazar
B
Becerril
D000595:Amino Acid Sequence; D057134:Antibodies, Neutralizing / Q000737:chemistry*; D000997:Antivenins / Q000096:biosynthesis; D061505:Cell Surface Display Techniques; D019020:Directed Molecular Evolution; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D008800:Mexico; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry*; D012604:Scorpion Venoms / Q000737:chemistry / Q000276:immunology*; D016415:Sequence Alignment; D020349:Surface Plasmon Resonance
D057134:Antibodies, Neutralizing; D000997:Antivenins; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; D012604:Scorpion Venoms
C. limpidus; C. noxius; C. suffusus; CDR; Centruroides limpidus limpidus Karsh; Centruroides noxius Hoffman; Centruroides suffusus suffusus; Cll1 and Cll2; Cn2 and Cn3; Css2; FW; Human scFv; In vitro maturation; Scorpion toxin; VH; VL; aa; amino acid(s); complementarity determining region (s); framework(s); scFv; single chain antibody fragment; toxin 2 of Centruroides suffusus suffusus; toxins 1 and 2 from Centruroides limpidus limpidus venom; toxins 2 and 3 from Centruroides noxius venom; variable domain of heavy chain; variable domain of light chain