Induction of neutralizing antibodies against Tityus serrulatus toxins by immunization with a recombinant nontoxic protein.
An immunogenic nontoxic protein (TsNTxP) was purified from the venom of the scorpion Tityus serrulatus (Ts). This peptide is composed of 63 amino acid residues with a high degree of structural homology with the toxins isolated from Ts. The nucleotide sequence of the gene that encodes TsNTxP was obtained and also showed a high degree of similarity with genes encoding Tityus toxins [Guatimosim, S.C.F., Prado, V.F., Diniz, C.R., Chávez-Olórtegui, C.. Kalapothakis, E., 1999. Molecular cloning and genomic analysis of TsNTxP; an immunogenic protein from Tityus serrulatus scorpion venom. Toxicon 37, 507-517]. In the present study the TsNTxP gene was expressed in E. coli BL21DE3 cells as a fusion protein with maltose-binding protein. The recombinant protein (TsNTxPrec) was purified by affinity chromatography and used as an immunogen in rabbits. The antigenic specificity of anti-TsNTxPrec antibodies was compared by an enzyme-linked immunosorbent assay using TsNTxP, TstFG50 (the fraction of Ts venom that represents most of the toxicity of the crude venom) and the crude venom, to coat microtitration plates. Anti-TsNTxPrec antibodies had a comparable high cross-reactivity for all antigens tested. Concentrations of Ts venom equivalent to 20 LD50 were effectively neutralized by 1 ml of the anti-TsNTxPrec serum. This result provides basic data for the use of such recombinant scorpion protein as an immunogen in the development of antivenoms for clinical use.
Guatimosim
S C
SC
Departamento de Bioquímica e Imunologia, ICB, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Kalapothakis
E
E
Diniz
C R
CR
Chávez-Olórtegui
C
C
D000818:Animals; D000997:Antivenins / Q000096:biosynthesis* / Q000302:isolation & purification; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D005822:Genetic Vectors; D007114:Immunization; D051379:Mice; D009004:Monosaccharide Transport Proteins / Q000276:immunology*; D009498:Neurotoxins / Q000276:immunology* / Q000633:toxicity; D009500:Neutralization Tests; D011817:Rabbits / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D012604:Scorpion Venoms / Q000276:immunology* / Q000633:toxicity; D014614:Vaccines, Synthetic / Q000276:immunology* / Q000302:isolation & purification
D000997:Antivenins; D009004:Monosaccharide Transport Proteins; C404939:NTxPrec protein, Tityus serrulat; D009498:Neurotoxins; D011994:Recombinant Proteins; D012604:Scorpion Venoms; D014614:Vaccines, Synthetic; C003609:tityustoxin