A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMS

A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMS

Phospholipase A2 (PLA2), a component of most snake venom toxins, cleaves 3-sn-phosphoglycerides releasing lysophosphatidyl-choline. The indirect quantitative assay method for PLA2 was standardized for specific antivenom titration in a fast and sensitive assay by the similarity with the hemolysis induced by PLA2 and by complement system in sheep erythrocytes. The curves obtained by plotting the degree of hemolysis against the doses of snake venom are concave to the abscissa axis following an equation similar to that previously described for the hemolysis induced by the C system. We observed that venoms of some Bothrops, Crotalus and Micrurus species contained around 1 x 10<img SRC="http:/img/fbpe/jvat/v2n2/image603.gif"> to 10<img SRC="http:/img/fbpe/jvat/v2n2/image604.gif"> Z/mg of venom, while the venom of Naja contained over one million Z/mg. Antibodies against PLA2 were titrated by incubating amounts of venom predetermined to give 1 to 5 Z with various dilutions of the antivenoms, and the remaining active PLA2 was determined in the hemolytic assay. We observed the following: a) the antivenoms contained specific antibodies against the PLA2 present in the corresponding venoms; b) cross-reactivity was not detected among PLA2 epitopes from venoms and nonspecific antivenoms; and c) the assay quantitatively performed determined the specific antibodies directed to epitopes on the molecule of PLA2. The method described in this paper is highly specific, sensitive and reproducible, besides being fast and inexpensive.

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Main Authors: ROCHA CAMPOS,A. C. M., FREITAS,M. C. W. DE, YAMAGUCHI,I. K., MARCELINO,J. R., DIAS DA SILVA,W.
Format: Digital revista
Language:English
Published: Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP, Universidade Estadual Paulista - UNESP 1996
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-79301996000200004
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spelling oai:scielo:S0104-793019960002000041999-01-11A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMSROCHA CAMPOS,A. C. M.FREITAS,M. C. W. DEYAMAGUCHI,I. K.MARCELINO,J. R.DIAS DA SILVA,W. PLA2 snake venoms immunoassay antivenoms indirect hemolysis Phospholipase A2 (PLA2), a component of most snake venom toxins, cleaves 3-sn-phosphoglycerides releasing lysophosphatidyl-choline. The indirect quantitative assay method for PLA2 was standardized for specific antivenom titration in a fast and sensitive assay by the similarity with the hemolysis induced by PLA2 and by complement system in sheep erythrocytes. The curves obtained by plotting the degree of hemolysis against the doses of snake venom are concave to the abscissa axis following an equation similar to that previously described for the hemolysis induced by the C system. We observed that venoms of some Bothrops, Crotalus and Micrurus species contained around 1 x 10<img SRC="http:/img/fbpe/jvat/v2n2/image603.gif"> to 10<img SRC="http:/img/fbpe/jvat/v2n2/image604.gif"> Z/mg of venom, while the venom of Naja contained over one million Z/mg. Antibodies against PLA2 were titrated by incubating amounts of venom predetermined to give 1 to 5 Z with various dilutions of the antivenoms, and the remaining active PLA2 was determined in the hemolytic assay. We observed the following: a) the antivenoms contained specific antibodies against the PLA2 present in the corresponding venoms; b) cross-reactivity was not detected among PLA2 epitopes from venoms and nonspecific antivenoms; and c) the assay quantitatively performed determined the specific antibodies directed to epitopes on the molecule of PLA2. The method described in this paper is highly specific, sensitive and reproducible, besides being fast and inexpensive.info:eu-repo/semantics/openAccessCentro de Estudos de Venenos e Animais Peçonhentos - CEVAP, Universidade Estadual Paulista - UNESPJournal of Venomous Animals and Toxins v.2 n.2 19961996-01-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-79301996000200004en10.1590/S0104-79301996000200004
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country Brasil
countrycode BR
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libraryname SciELO
language English
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author ROCHA CAMPOS,A. C. M.
FREITAS,M. C. W. DE
YAMAGUCHI,I. K.
MARCELINO,J. R.
DIAS DA SILVA,W.
spellingShingle ROCHA CAMPOS,A. C. M.
FREITAS,M. C. W. DE
YAMAGUCHI,I. K.
MARCELINO,J. R.
DIAS DA SILVA,W.
A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMS
author_facet ROCHA CAMPOS,A. C. M.
FREITAS,M. C. W. DE
YAMAGUCHI,I. K.
MARCELINO,J. R.
DIAS DA SILVA,W.
author_sort ROCHA CAMPOS,A. C. M.
title A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMS
title_short A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMS
title_full A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMS
title_fullStr A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMS
title_full_unstemmed A SENSITIVE AND SPECIFIC IMMUNOASSAY FOR THE MEASUREMENT OF THE ANTIBODIES PRESENT IN HORSE ANTIVENOMS ENDOWED WITH THE CAPACITY TO BLOCK THE PHOSPHOLIPASE A2-DEPENDENT HEMOLYSIS INDUCED BY SNAKE VENOMS
title_sort sensitive and specific immunoassay for the measurement of the antibodies present in horse antivenoms endowed with the capacity to block the phospholipase a2-dependent hemolysis induced by snake venoms
description Phospholipase A2 (PLA2), a component of most snake venom toxins, cleaves 3-sn-phosphoglycerides releasing lysophosphatidyl-choline. The indirect quantitative assay method for PLA2 was standardized for specific antivenom titration in a fast and sensitive assay by the similarity with the hemolysis induced by PLA2 and by complement system in sheep erythrocytes. The curves obtained by plotting the degree of hemolysis against the doses of snake venom are concave to the abscissa axis following an equation similar to that previously described for the hemolysis induced by the C system. We observed that venoms of some Bothrops, Crotalus and Micrurus species contained around 1 x 10<img SRC="http:/img/fbpe/jvat/v2n2/image603.gif"> to 10<img SRC="http:/img/fbpe/jvat/v2n2/image604.gif"> Z/mg of venom, while the venom of Naja contained over one million Z/mg. Antibodies against PLA2 were titrated by incubating amounts of venom predetermined to give 1 to 5 Z with various dilutions of the antivenoms, and the remaining active PLA2 was determined in the hemolytic assay. We observed the following: a) the antivenoms contained specific antibodies against the PLA2 present in the corresponding venoms; b) cross-reactivity was not detected among PLA2 epitopes from venoms and nonspecific antivenoms; and c) the assay quantitatively performed determined the specific antibodies directed to epitopes on the molecule of PLA2. The method described in this paper is highly specific, sensitive and reproducible, besides being fast and inexpensive.
publisher Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP, Universidade Estadual Paulista - UNESP
publishDate 1996
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-79301996000200004
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