High sensitivity ELISA for detection of botulinum neurotoxin serotype F.
Botulinum neurotoxins (BoNTs) are classified as category A biological threat agents by the Centers for Disease Control and Prevention (CDC) in the United States for its hazardous and potential bioterrorist threat to the public. About 1% naturally occurring botulisms are caused by Botulinum neurotoxin serotype F (BoNT/F). Most of the immunoassays for detecting BoNTs focus on the serotypes A and B, but few methods have been established for the detection of BoNT/F. Recently, the recombinant Hc subunit of botulinum neurotoxin type F (rFHc) was expressed as an effective vaccine against BoNT/F, indicating that this rFHc could be an effective immunogen to raise monoclonal antibodies (MAbs) for the detection and neutralization of BoNT/F. Here we present a novel sandwich enzyme-linked immunosorbent assay (ELISA) based on two MAbs against rFHc, which were FMMU-BTF-8 and FMMU-BTF-29 as capture antibody and detection antibody, respectively. The limit of detection (LOD) of this ELISA reached 12.09 pg/mL, much less than that of the other reported immunoassays. A simple, sensitive ELISA for detecting and quantifying BoNT/F was established, which can be used as a valuable method to detect and quantify BoNT/F.
Department of Immunology, The Fourth Military Medical University, Xi'an, Shaanxi Province, People's Republic of China.
Zhijia
Z
Liu
Chunmei
C
Zhang
Yongming
Y
Li
Chaojun
C
Song
Yuanjie
Y
Sun
Yuying
Y
Wei
Zhuwei
Z
Xu
Angang
A
Yang
Zhikai
Z
Xu
Kun
K
Yang
Boquan
B
Jin
D000818:Animals; D055910:Antibodies, Immobilized / Q000096:biosynthesis / Q000737:chemistry*; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis / Q000737:chemistry*; D001905:Botulinum Toxins / Q000276:immunology*; D002138:Calibration; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D005260:Female; D005516:Food Microbiology; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry*; D057230:Limit of Detection; D008460:Meat / Q000032:analysis / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008892:Milk / Q000737:chemistry / Q000382:microbiology; D011485:Protein Binding; D012015:Reference Standards; D015203:Reproducibility of Results
D055910:Antibodies, Immobilized; D058846:Antibodies, Monoclonal, Murine-Derived; D007074:Immunoglobulin G; D001905:Botulinum Toxins; C096321:botulinum toxin type F