RRC ID 44592
Author Hara Y, Dong J, Ueda H.
Title Open-sandwich immunoassay for sensitive and broad-range detection of a shellfish toxin gonyautoxin.
Journal Anal Chim Acta
Abstract At present, the analytical method for paralytic shellfish poisoning (PSP) toxins in shellfish is the mouse bioassay (MBA), which is an official method of the Association of Analytical Communities (AOAC [8]). However, the low sensitivity and concerns over the number of live animals required for testing have been cited as the major reason for seeking its replacement. In this report, we employed an open-sandwich immunoassay (OS-IA) to detect gonyautoxin (GTX2/3), a kind of PSP toxins. OS-IA, which utilizes the antigen-induced enhancement of antibody VH/VL interaction, can measure a small molecule antigen in a noncompetitive format. Hence it has a wider working range and shorter measurement time. We isolated anti-GTX2/3 antibody gene from a hybridoma GT-13A by screening a Fab-displaying phage library. Then the vectors for OS-IA were constructed, and examined for antigen concentration-dependency of the VH/VL interaction by OS-ELISA. As a result, in each case, signal intensity increases notably in a wide concentration range (0.1 to >1000 ng mL(-1)) of free GTX2/3, which was enough to cover its regulation value (80 μg 100 g(-1)) in many countries. So OS-IA will be widely applicable to detect PSP toxins in shellfish meats and in drinking water.
Volume 793
Pages 107-13
Published 2013-9-2
DOI 10.1016/j.aca.2013.07.024
PII S0003-2670(13)00970-7
PMID 23953213
MeSH Amino Acid Sequence Animals Antibodies / genetics Antibodies / immunology Drinking Water / analysis Enzyme-Linked Immunosorbent Assay* Immunoglobulin Variable Region / chemistry Molecular Sequence Data Peptide Library Saxitoxin / analogs & derivatives* Saxitoxin / analysis Shellfish / analysis*
IF 5.977
Times Cited 15
Human and Animal Cells