RESUMO
Using a panel of monoclonal antibodies (mAb) against human myoglobin (Mb), we have shown that the sensitivity of antigen-capture enzyme-linked immunosorbent assay (ELISA) may be significantly increased by the simultaneous immobilization on a solid phase of two co-operating capture mAbs. This method ("a three-site ELISA") uses three mAbs at different epitopes of the same antigen (two capture/one tracer), unlike the traditional two-site assay, using one capture and one tracer mAbs. We established two-site and three-site ELISA assays for Mb, by varying capture and tracer mAbs. Three-site assays showed 4-6 fold increase in sensitivity, if compared with two-site assays. The model for the effect has been suggested, according to which in three-site ELISA the high-affinity cyclic configurations may be formed by an antigen, two-capture mAbs and the surface of solid phase.
Assuntos
Anticorpos Monoclonais/química , Antígenos/química , Ensaio de Imunoadsorção Enzimática/métodos , Epitopos/química , Animais , Especificidade de Anticorpos , Calibragem , Células Cultivadas , Cromatografia de Afinidade , Cromatografia por Troca Iônica , Peroxidase do Rábano Silvestre/química , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Miocárdio/química , Mioglobina/imunologia , Proteínas/química , Coelhos , Padrões de ReferênciaRESUMO
A new operation to eliminate accessory pathways--epicardial electrical ablation--is described. In a group of 201 patients without concomitant disease, the mortality rate was 0.5% and the overall efficacy of the operation for free wall accessory pathways, 98%. A retrospective clinical study of 44 unselected patients was performed to examine how safe epicardial electrical ablation is. The criteria for intraoperative effectiveness were disappearance of both the delta wave and retrograde conduction and inability to induce tachycardia. In the postoperative and follow-up periods, the following were reviewed: electrocardiograms; Holter monitor recordings (24 to 26 hours); release of the myocardial-specific isoenzyme of creatine kinase; intracardiac hemodynamics and myocardial contractility (radionuclide methods); selective coronary arteriograms and ventriculograms; mean work capacity (bicycle ergometer); diagnostic transesophageal electrical stimulation; and histology of the area of ablation. The main conclusion of this study is that epicardial electrical ablation is a highly efficient and safe operation for surgical elimination of parietal accessory pathways in patients with Wolff-Parkinson-White syndrome. Its advantages are its technical simplicity and the opportunity to review results immediately during the operation.