Centro de Ciências da Saúde

URI Permanente desta comunidadehttps://repositorio.ufrn.br/handle/1/21

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Development of immunoassays for anti-electronegative LDL autoantibodies and immune complexes
(Clinica Chimica Acta, 2012-01) Evangelista, Karine Cavalcanti Maurício Sena; Faulin, Tanize do Espirito Santo; Pacheco, Débora Bezerra; Augusto, Elaine Moura; Abdalla, Dulcineia Saes Parra
Background: Electronegative low-density lipoprotein (LDL−) promotes atherosclerosis through inflammatory and immunologic mechanisms that lead to the production of anti-LDL(−) autoantibodies and to the subsequent formation of immune complexes (IC) and macrophage foam cells. We described the development and validation of an ELISA for the quantification of free anti-LDL(−) autoantibodies and an ELISA for the quantification of IC consisting of LDL(−)-bound IgG in human plasma. Methods: LDL(−) purified from human plasma, and anti-LDL(−) monoclonal antibody Fab fragments were adsorbed onto ELISA plates to capture anti-LDL(−) autoantibodies and IC-LDL(−), respectively. The performance characteristics of both ELISAs, including the limits of detection and quantification, accuracy and interand intra-assay precision were evaluated. Linearity, interference and stability tests were also performed. Results: The calibration range of the anti-LDL(−) assay was 0.004–0.125 mU/l and plasma demonstrated a dilutional linearity when diluted 1:100, 1:200, 1:400 and 1:800. The calibration range of the IC-LDL(−) assay was 0.06–4 U/l, and plasma demonstrated a dilutional linearity when diluted 1:12.5, 1:25, 1:50 and 1:100. Both ELISAs showed intra- and inter-assay precision and recovery within the required limits for immunoassays. Conclusion: These ELISAs can be used in clinical studies and for the biochemical investigation of atherosclerosis. In addition, they will enable the comprehensive evaluation of the importance of bound or free autoantibodies against LDL(−) in this disease
Validation of a novel ELISA for measurement of electronegative low-density lipoprotein
(Clinical Chemistry and Laboratory Medicine, 2008-01) Evangelista, Karine Cavalcanti Maurício Sena; Faulin, Tanize do Espírito Santo; Telles, Andréia Elisa Rodrigues; Grosso, Daniela de Mattos; Faulin, Edson José Bernardi; Abdalla, Dulcineia Saes Parra
Background: Oxidative modification of low-density lipoprotein (LDL) plays a key role in the pathogenesis of atherosclerosis. LDL(–) is present in blood plasma of healthy subjects and at higher concentrations in diseases with high cardiovascular risk, such as familial hypercholesterolemia or diabetes. Methods: We developed and validated a sandwich ELISA for LDL(–) in human plasma using two monoclonal antibodies against LDL(–) that do not bind to native LDL, extensively copper-oxidized LDL or malondialdehyde-modified LDL. The characteristics of assay performance, such as limits of detection and quantification, accuracy, inter- and intra-assay precision were evaluated. The linearity, interferences and stability tests were also performed. Results: The calibration range of the assay is 0.625–20.0 mU/L at 1:2000 sample dilution. ELISA validation showed intra- and inter-assay precision and recovery within the required limits for immunoassays. The limits of detection and quantification were 0.423 mU/L and 0.517 mU/L LDL(–), respectively. The intra- and inter-assay coefficient of variation ranged from 9.5% to 11.5% and from 11.3% to 18.9%, respectively. Recovery of LDL(–) ranged from 92.8% to 105.1%. Conclusions: This ELISA represents a very practical tool for measuring LDL(–) in human blood for widespread research and clinical sample use

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Navegando Centro de Ciências da Saúde por Autor "Abdalla, Dulcineia Saes Parra"