Title
Production of monoclonal antibodies capable of neutralizing dermonecrotic activity of Loxosceles intermedia spider venom and their use in a specific immunometric assay
Date Issued
01 January 2003
Access level
metadata only access
Resource Type
journal article
Author(s)
Alvarenga L.M.
Martins M.S.
Moura J.F.
Kalapothakis E.
Oliveira J.C.
Mangili O.C.
Granier C.
Universidade Federal de Minas Gerais
Abstract
We have produced 13 mAbs for Loxosceles intermedia crude venom. Twelve were reactive against proteins of 32-35 kDa and one of these Li mAb(7) showed high neutralizing potency for the dermonecrotic activity of L. intermedia venom. This Li mAb(7) showed no cross-reactivity, with Loxosceles laeta (Brazil), L. laeta (Perú) and Loxosceles gaucho venoms. The mAbs were produced by immunization with the crude venom and screened by enzyme-linked immunosorbent assay (ELISA) using L. intermedia whole venom or dermonecrotic fraction (DNF) as antigens coated onto microtitre plates. A sensitive two-site immunometric assay was designed and shown to be useful for identifying and quantifying DNF from L. intermedia in biological samples. The Li mAb(7) coated onto microtitre plates and hyperimmune horse anti-L. intermedia IgGs prepared by immunoaffinity chromatography and conjugated to horseradish peroxidase were used to set up a sandwich-type ELISA. Measurable absorbance signals were obtained with 0.2 ng of L. intermedia crude venom per assay. © 2003 Elsevier Ltd. All rights reserved.
Start page
725
End page
731
Volume
42
Issue
7
Language
English
OCDE Knowledge area
Toxicología
Subjects
Scopus EID
2-s2.0-1642452868
PubMed ID
Source
Toxicon
ISSN of the container
00410101
Sponsor(s)
Funding text
We thank Dr Michael Richardson for critical comments and editorial assistance. This research was supported by grants from FAPEMIG, Contract N° CBB 2334/97 and by CNPq, Contracts N° 479643/01-6 and 690119/01-1 Bilateral Program funded by the Institut National de la Santé et de la Recherche Médicale-INSERM; France and CNPq, Brazil. C. Chávez-Olórtegui and E. Kalapothakis are recipients of CNPq fellowships.
Sources of information:
Directorio de Producción Científica
Scopus