The co-purification of a lectin (BJcuL) with phospholipases A2 from Bothrops jararacussu snake venom by immunoaffinity chromatography with antibodies to crotoxin.
Antigens of Bothrops jararacussu snake venom cross-reacting with specific antibodies against crotoxin, an Asp49 PLA(2)-containing heterodimeric complex from Crotalus durissus terrificus snake venom, were purified by two steps of immunoaffinity chromatography. The resulting fraction (Bj-F) was shown to be non-toxic (to mice and rabbits) and immunogenic to rabbits. Antibodies raised against Bj-F were able to protect mice against the lethal effect of both B. jararacussu and Crotalus durissus terrificus snake venoms. Then, the procedure developed showed to be useful for the rapid preparation of an antigen able to elicit neutralizing antibodies against the lethal activities of both venoms. Further fractionation of Bj-F revealed the concomitant presence of two major components: BJcuL, a lectin present in B. jararacussu venom, and BthTX-I, a Lys49 PLA(2) homolog, besides other molecules in minor amounts. Our data are discussed and raise the point that the presence of unrelated molecules may be taken into account when immuno-based methods are considered for purification purposes.
Gomes
Paulo César
PC
Fundação Ezequiel Dias (FUNED), Rua Conde Pereira Carneiro 80, CEP 30510-010, Belo Horizonte, MG, Brazil.
Machado de Avila
Ricardo A
RA
Selena Maria
Wany
W
Richardson
Michael
M
Fortes-Dias
Consuelo Latorre
CL
Chávez-Olórtegui
Carlos
C
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000997:Antivenins / Q000276:immunology*; D015153:Blotting, Western; D017837:Bothrops; D005591:Chemical Fractionation; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003439:Crotoxin / Q000737:chemistry* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D037102:Lectins / Q000032:analysis / Q000302:isolation & purification*; D007928:Lethal Dose 50; D051379:Mice; D054467:Phospholipases A2 / Q000302:isolation & purification*; D011817:Rabbits
D000906:Antibodies; D000997:Antivenins; D037102:Lectins; D003439:Crotoxin; D054467:Phospholipases A2