Examinando por Materia "Antibodies, Monoclonal"

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Acceso abierto
Optimal parameters to standardize a sandwich type Elisa with monoclonal antibodies for the diagnosis of human neurocysticercosis in urine samples.
(Universidad Privada Norbert Wiener, 2020-02-22) Santos Salazar, Yesica; Benites Azabache, Juan Carlos
Human neurocysticercosis (NCC) is an infection caused by the cysticercus of T. solium, which primarily affects the brain, mostly causing epilepsy in developing countries and being a public health problem. A variety of serological techniques have been implemented and adapted to improve the diagnosis and study of this parasitosis; however, there are few studies using monoclonal antibodies in their development. The aim of this study was to determine the optimal parameters to standardize a sandwich-type ELISA using monoclonal anti-T. solium antibodies in urine samples. The biotinylation of the 4 mAbs (TsW5, TsW7, TsW8, and TsW11) was successfully carried out, and during simultaneous plate selection, Immulón 4HBX was determined as the best option, with a blocking solution of 2% Carnation milk showing the best ratio of 23.6. The adherence buffer was selected as Dulbecco’s phosphate-buffered saline (PBS) pH 7.2 (Oxoid), determining the optimal concentration for the best pair of mAbs for antigen detection, resulting in the capture antibody TsW8 (4 µg/ml) and the biotinylated detection antibody TsW5 (4 µg/ml). Among substrate options, O-Phenylene diamine dihydro-Chloride (OPD) presented the highest ratio (64) with a 15-minute incubation time, followed by the stop solution (H2SO4 2N). All evaluations were performed with positive and negative urine control samples, with reactivity expressed as a ratio (positive OD / negative OD). The optimal parameters for the designed sandwich-type ELISA show its potential utility for diagnosis and may be useful for post-treatment monitoring of patients with viable NCC infections.