A gold nanoparticle-single-chain fragment variable antibody as an immunoprobe for rapid detection of morphine by dipstick

Gandhi, S.; Banga, I.; Maurya, P.K.; Eremin, S.A.

Авторы: Gandhi Sonu, Banga Ivneet, Maurya Pawan Kumar, Eremin Sergei A.
Журнал: RSC advances
Том: 8
Номер: 3
Год издания: 2018
Издательство: Royal Society of Chemistry
Местоположение издательства: United Kingdom
Первая страница: 1511
Последняя страница: 1518
DOI: 10.1039/c7ra12810j
Аннотация: Gold nanoparticle (AuNP)-based optical assays are of significant interest since the molecular phenomenon can be examined easily with change in the color of AuNPs. Herein, we report the development of a dipstick using a AuNP-labeled single-chain fragment variable (scFv) antibody for the detection of morphine. The scFv antibodies for morphine were developed using phage display-based antibody library. Immunoglobulin variable regions of heavy (VH)- and light (VL)-chain genes were connected via a glycine–serine linker isolated from murine immune repertoire and cloned into the expression vector pIT2. The scFv was produced in Escherichia coli HB2151, yielding a functional protein with a molecular weight of approximately 32 kDa. The morphine scFv was labeled with gold nanoparticles and used as an optical immunoprobe in a dipstick. The competitive dipstick assay characterized the ability of the scFv antibody to recognize free morphine. The detection range was 1–1000 ng mL_1 with a limit of detection (LOD) of 5 ng mL_1 under optimal conditions, and the IC50 value was 14 ng mL_1 for morphine. The developed optical dipstick kit of scFv antibody was capable of specifically binding to free morphine and its analogs in a solution in less than 5 min and could be useful for on-site screening of a real sample in blood, urine, and saliva.
Добавил в систему: Еремин Сергей Александрович

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A gold nanoparticle-single-chain fragment variable antibody as an immunoprobe for rapid detection of morphine by dipstickстатья

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