RESUMO
Neurocysticercosis is a parasitic disease of major public health importance. Definitive diagnosis requires neuroimaging, which is typically unavailable in rural impoverished regions of endemicity. Screening immunoassays can support diagnosis in this setting by identifying individuals most likely to have severe forms of disease for referral to imaging. Urine sampling is convenient, painless, and generally well accepted. We developed a rapid point-of-care (POC) assay to detect urinary antigens and assessed concordance with a standard antigen ELISA (Ag-ELISA), both using monoclonal antibodies TsW8/TsW5. From 28,145 stored community samples with Ag-ELISA results, we selected 843 for comparison, 281 each from nonreactive (ratio <1), reactive-below-cutoff (ratio 1:3), and positive (ratio ≥3) samples. Overall agreement was 73.6%, with strong agreement observed in the nonreactive (280/281, 99.6%) and positive (255/281, 90.8%) groups. This affordable noninvasive POC test can be applied to identify individuals in the community most at risk of developing severe disease.
RESUMO
RESUMEN Se evaluó el uso de partículas magnéticas acopladas a proteína L para la concentración y purificación de anticuerpos monoclonales inmunoglobulina M (mIgM) contra Taenia solium. Se evaluaron tres métodos de concentración y diferentes tiempos de elución y se optimizó la proporción de partículas a la proporción de mIgM. Demostramos que: 1) con el uso partículas magnéticas no se requiere de una concentración previa de mIgM, lo que disminuye la manipulación de los anticuerpos y mejora la recuperación, 2) se puede omitir el uso de un tampón de unión, ya que el pH de la mayoría de los sobrenadantes de cultivo celular son neutros, y 3) se necesitan tiempos de elución más largos (~45 minutos) para aumentar la recuperación a un nivel mayor a 80%. El estudio demuestra que el uso de partículas magnéticas acopladas a proteína L es una herramienta simple y eficiente para la concentración y purificación de mIgM.
ABSTRACT The use of L protein coupled magnetic particles for the concentration and purification of immunoglobulin M (mIgM) monoclonal antibodies against Taenia solium was evaluated. Three concentration methods and different elution times were evaluated and the ratio of particles to the ratio of mIgM was optimized. It is demonstrated that: 1) with the use of magnetic particles, a previous concentration of mIgM is not required, which reduces the manipulation of the antibodies and improves the recovery, 2) the use of a binding buffer can be omitted, since the pH of most cell culture supernatants are neutral, and 3) longer elution times (~ 45 minutes) are needed to increase recovery to a level greater than 80%. The study demonstrates that the use of L protein-coupled magnetic particles is a simple and efficient tool for mIgM concentration and purification.