Green Fluorescent Protein-labeled Recombinant Fluobody for Detecting the Picloram Herbicide

Kim, In Soo; Shim, Jae Han; Suh, Yong Tack; Yau, Kerrm; Hall, J. Christopher; Trevors, J. T.; Lee, Hung

doi:10.1271/bbb.66.1148

Cited by 15 publications

(4 citation statements)

References 12 publications

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“…In addition, sensitive FLISA can be performed by taking advantage of the strong fluorescence intensity of GFP [48]. Up to date, ic-FLISA can be developed to detect/determine small molecules, which include picloram [49], s-triazine [50], 5-methyl 2'-deoxycytidine [51] and bioactive natural product, plumbagin (PL) and ginsenoside Re (GRe) as previously described in our group [52]- [54]. The icFLISA for PL and GRe shows a lower limit of determination than conventional icELISA.…”

Section: Fluorescence-linked Immunosorbent Assaymentioning

confidence: 99%

Application of Green Fluorescent Protein in Immunoassays

Sakamoto¹,

Shoyama²,

Tanaka³

et al. 2014

ABB

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Green fluorescent protein (GFP) is a protein that emits green fluorescence when exposed to a radiation of ultraviolet wavelength range, even without the addition of substrate and cofactor. Because of such characteristics, the usage of GFP is widespread in both in vivo and in vitro applications. In addition, recent advances in biotechnology have enabled GFP to be expressed in various hosts, including bacteria, yeast, plants, animals, and even living-cells, for multiple purposes. Currently, GFP is a subject of great interest in the analytical sciences, especially in immunoassays for qualitative and quantitative analyses, when it is fused with an antibody because of the high sensitivity of GFP and antigen-binding specificity of antibodies. Recently the fluobody, which is a fusion protein of GFP with single-chain variable fragment antibody (scFv), has become a useful tool in various fields. We review here the applications of GFP as fluobodies in immunoassays.

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Section: Fluorescence-linked Immunosorbent Assaymentioning

confidence: 99%

Application of Green Fluorescent Protein in Immunoassays

Sakamoto¹,

Shoyama²,

Tanaka³

et al. 2014

ABB

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“…Though these methods are very sensitive and have lower LODs, they usually need expertise, lengthy sample preparation, purification processes, and sophisticated laboratory setups. Although radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA) have shown to be efficient techniques for picloram detection, their use for on-site pesticide management is constrained by their complex washing procedures, lengthy incubation times, radiation risks, and huge instruments 18 , 19 . Therefore, a more practical, simple, and cost-effective approach is required.…”

Section: Introductionmentioning

confidence: 99%

Chemically engineered unzipped multiwalled carbon nanotube and rGO nanohybrid for ultrasensitive picloram detection in rice water and soil samples

Dkhar

Kumari

Chandra

2023

Sci Rep

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Picloram (4-Amino-3,5,6-trichloro pyridine-2-carboxylic acid) is a chlorinated herbicide that has been discovered to be tenacious and relatively durable in both soil and water. It is known to have adverse and unpleasant effects on humans causing several health complications. Therefore, the determination of picloram is profoundly effective because of its bio-accumulative and persistent nature. Because of this, a sensitive, rapid, and robust detection system is essential to detect traces of this molecule. In this study, we have constructed a novel nanohybrid system comprising of an UZMWCNT and rGO decorated on AuNPs modified glassy carbon electrode (UZMWCNT + rGO/AuNPs/GCE). The synthesized nanomaterials and the developed system were characterized using techniques such as SEM, XRD, SWV, LSV, EIS, and chronoamperometry. The engineered sensor surface showed a broad linear range of 5 × 10–2 nM to 6 × 105 nM , a low limit of detection (LOD) of 2.31 ± 0.02 (RSD < 4.1%) pM and a limit of quantification (LOQ) of 7.63 ± 0.03 pM. The response time was recorded to be 0.2 s, and the efficacy of the proposed sensor system was studied using rice water and soil samples collected from the agricultural field post filtration. The calculated recovery % for picloram in rice water was found to be 88.58%—96.70% (RSD < 3.5%, n = 3) and for soil it was found to be 89.57%—93.24% (RSD < 3.5%, n = 3). In addition, the SWV responses of both the real samples have been performed and a linear plot have been obtained with a correlation coefficient of 0.97 and 0.96 for rice and soil samples, respectively. The interference studies due to the coexisting molecules that may be present in the samples have been found to be negligible. Also, the designed sensor has been evaluated for stability and found to be highly reproducible and stable towards picloram detection.

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“…Currently, the usage of fluobody has been expanded from diagnosis, molecular targeted therapy, immunolabeling of cancer cells, probes for immunoassay, fluorescence-activated cell sorter (FACS) to fluorescent-linked immunosorbent assay (FLISA) for the detection of large molecular weight antigens such as proteins, peptides, and microtubules [47][48][49][50][51]. So far the application of a fluobody in FLISA for the detection of small molecules has been rarely reported except against herbicide, picloram [52] and s-triazine [53], and bioactive natural products, PL and G-Re [54][55][56] previously described by our group.…”

Section: Introductionmentioning

confidence: 99%

Fluobodies against Bioactive Natural Products and their Application in Fluorescence-Linked Immunosorbent Assay

et al. 2012

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An enzyme-linked immunosorbent assay (ELISA) using monoclonal antibody (MAb), Fab antibody, and single-chain variable fragment (scFv) antibody has become one of the most promising analytical methods owing to its rapidity, sensitivity, and reliability. Recently, a chimera of green fluorescent protein (GFP) with a scFv antibody, named fluobody, was proposed as a probe for an alternative immunosorbent assay; i.e., fluorescence-linked immunosorbent assay (FLISA). In this FLISA, an even more sensitive, simple, and rapid immunoassay can be performed by detecting the highly sensitive fluorophore of GFP that is genetically and directly fused to the scFv antibody. In addition, the time- and cost-consuming secondary antibody reaction and the following enzyme-substrate reaction, necessary for conventional ELISA, can be avoided, making it possible to complete the assay more rapidly. Focusing on naturally occurring bioactive products, fluobody recognizing 1,4-naphthoquinone, plumbagin and triterpenoid saponin, ginsenosides were successfully expressed in Escherichia coli (E. coli) and applied to FLISA. The construction, the expression, and the potential use of fluobody in quantitative/qualitative analysis of bioactive natural products are reviewed in this article

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Green Fluorescent Protein-labeled Recombinant Fluobody for Detecting the Picloram Herbicide

Cited by 15 publications

References 12 publications

Application of Green Fluorescent Protein in Immunoassays

Application of Green Fluorescent Protein in Immunoassays

Chemically engineered unzipped multiwalled carbon nanotube and rGO nanohybrid for ultrasensitive picloram detection in rice water and soil samples

Fluobodies against Bioactive Natural Products and their Application in Fluorescence-Linked Immunosorbent Assay

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