Сэндвичи, картошка, немного антигенов…и флагманский Malvern Zetasizer – рецепт блестящей научной статьи

Очередная высококлассная и интересная работа от одной из ведущих лабораторий Института биохимии имени А. Н. Баха РАН (ИНБИ РАН) — Лаборатории иммунобиохимии…

Статья: How to Improve Sensitivity of Sandwich Lateral Flow Immunoassay for Corpuscular Antigens on the Example of Potato Virus Y?

Авторы: Shyatesa C. Razo, Vasily G. Panferov, Irina v. Safenkova, Yuri A. Varitsev, Anatoly v. Zherdev, Elena N. Pakina, and Boris B. Dzantiev

Опубликовано: Sensors (Basel). 2018 Nov; 18(11): 3975.

Аннотация

A simple approach was proposed to decrease the detection limit of sandwich lateral flow immunoassay (LFIA) by changing the conditions for binding between a polyvalent antigen and a conjugate of gold nanoparticles (GNPs) with antibodies. In this study, the potato virus Y (PVY) was used as the polyvalent antigen, which affects economically important plants in the Solanaceae family. The obtained polyclonal antibodies that are specific to PVY were characterized using a sandwich enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). For LFIA, the antibodies were conjugated with GNPs with a diameter of 17.4 ± 1.0 nm. We conducted LFIAs using GNP conjugates in a dried state on the test strip and after pre-incubation with a sample. Pre-incubating the GNP conjugates and sample for 30 s was found to decrease the detection limit by 60-fold from 330 ng∙mL−1 to 5.4 ng∙mL−1 in comparison with conventional LFIA. The developed method was successfully tested for its ability to detect PVY in infected and uninfected potato leaves. The quantitative results of the proposed LFIA with pre-incubation were confirmed by ELISA, and resulted in a correlation coefficient of 0.891. The proposed approach is rapid, simple, and preserves the main advantages of LFIA as a non-laboratory diagnostic method.

Keywords: gold nanoparticles, increase of sensitivity, lateral flow test strips, potato virus Y, pre-incubation, sandwich immunoassay

В исследовании применялись результаты измерений образцов на анализаторе Zetasizer.

Текст статьи для ознакомления доступен по ссылке: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6263755/