Development of process to produce polyvalent IgY antibodies anti-African snake venom.
Polyvalent anti-Bitis and anti-Naja antivenom IgY antibodies were prepared using B. arietans, B. nasicornis, B. rhinoceros, N. melanoleuca, and N. mossambica venoms to immunize chickens. Blood and eggs were collected before and during the 10-month immunization period; the sera and yolk extracts were then prepared and assayed for the presence of antivenom antibodies by ELISA and Western blot methods. ELISA Antivenom antibody titers, referred to as U-ELISA/ml of serum or egg yolk extracts, absent in pre-immunization sera or yolk, increased sharply during the 4 weeks after immunization, reaching a plateau thereafter. Yolk extracts with high antivenom titers, as detected by ELISA were used to isolate and purify IgY. Purified IgY preparations recognized venom protein bands from 10 to 20 kDa to 60 and 70 kDa, as shown by Western blot. Recovery of antivenom antibodies from the whole yolk was over 80%. Final preparations exhibited high antivenom activity (>100,000 U-ELISA/ml) as well as efficacy in neutralizing venom lethality (1,440 microg of IgY neutralize 62.2 LD(50) of venom), and were free of toxic products, pyrogen or bacterial and fungal contaminations.
de Almeida
Cláudia Maria Costa
CM
Laboratório de Sanidade Animal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense-Darcy Ribeiro, Campos dos Gottacazes, RJ, Brazil.
da Silva
Cláudia Letícia
CL
Couto
Humberto Pena
HP
Escocard
Rita de Cássia Mothé
Rde C
da Rocha
David Gitirana
DG
Sentinelli
Lynna de Paula
Lde P
Kipnis
Thereza Liberman
TL
da Silva
Wilmar Dias
WD
D000818:Animals; D021461:Animals, Outbred Strains; D000906:Antibodies / Q000276:immunology; D000997:Antivenins / Q000276:immunology*; D002645:Chickens / Q000276:immunology; D004530:Egg Yolk / Q000276:immunology; D004546:Elapid Venoms / Q000737:chemistry / Q000276:immunology*; D017815:Elapidae; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007136:Immunoglobulins / Q000276:immunology*; D007928:Lethal Dose 50; D051379:Mice; D009500:Neutralization Tests; D011817:Rabbits
D000906:Antibodies; D000997:Antivenins; D004546:Elapid Venoms; C016512:IgY; D007136:Immunoglobulins