Phytoproducts are involved in various fields of industry. Small-molecule (Mw < 900 Da) organic compounds can be used to indicate the quality of plant samples in the perspective of efficacy by ...measuring the necessary secondary metabolites and in the perspective of safety by measuring the adulterant level of toxic compounds. The development of reliable detection methods for these compounds in such a complicated matrix is challenging. The lateral flow immunoassay (LFA) is one of the immunoassays well-known for its simplicity, portability, and rapidity. In this review, the general principle, components, format, and application of the LFA for phytoproducts are discussed.
Abstract Atacama Large Millimeter/submillimeter Array (ALMA) data toward QSO J1851+0035 ( l = 33.°498, b = +0.°194) were used to study absorption lines by Galactic molecular gas. We detected 17 ...species (CO, 13 CO, C 18 O, HCO + , H 13 CO + , HCO, H 2 CO, C 2 H, c -C 3 H, c -C 3 H 2 , CN, HCN, HNC, CS, SO, SiO, and C) and set upper limits to 18 species as reference values for chemical models. About 20 independent velocity components at 4.7–10.9 kpc from the Galactic center were identified. Their column density and excitation temperature estimated from the absorption study, as well as the CO intensity distributions obtained from the FUGIN survey, indicate that the components with τ ≲1 correspond to diffuse clouds or cloud outer edges. Simultaneous multiple-Gaussian fitting of CO J = 1–0 and J = 2–1 absorption lines shows that these are composed of narrow- and broad-line components. The kinetic temperature empirically expected from the high HCN/HNC isomer ratio (≳4) reaches ≳40 K and the corresponding thermal width accounts for the line widths of the narrow-line components. CN-bearing molecules and hydrocarbons have tight and linear correlations within the groups. The CO/HCO + abundance ratio showed a dispersion as large as 3 orders of magnitude with a smaller ratio in a smaller N (HCO + ) (or lower A V ) range. Some of the velocity components are detected in single-dish CO emission and ALMA HCO + absorption but without corresponding ALMA CO absorption. This may be explained by the mixture of clumpy CO emitters not resolved with the ∼1 pc single-dish beam surrounded by extended components with a very low CO/HCO + abundance ratio (i.e., CO-poor gas).
The establishment of the Nobeyama Radio Observatory (NRO) in 1982 was an important event that greatly influenced the subsequent development of Japanese astronomy. The 45 m radio telescope and the ...Nobeyama Millimeter Array (NMA) pushed Japanese radio astronomy to the forefront of the world. As a plan beyond the Nobeyama telescopes, the Japanese radio astronomy community considered a large array to achieve unprecedented resolution at millimeter and submillimeter wavelengths under the project name of the Large Millimeter and Submillimeter Array (LMSA). After long and patient discussions and negotiations with the United States and Europe, the LMSA plan eventually led to the ALMA (Atacama Large Millimeter/submillimeter Array) as an international joint project, and the ALMA was inaugurated in 2013. This paper reviews the process from the establishment of the NRO to the realization of the ALMA, including planning of the LMSA, international negotiations, site survey, instrumental developments, and initial science results.
Licochalcone A (LicoA), which is found in the root of Chinese licorice (Glycyrrhiza inflata Batalin), has been reported as an effective anti-microbial and anti-inflammation and was approved for the ...treatment of rosacea and acne in clinical therapeutic. To develop a monoclonal antibody against LicoA, a mannich reaction hapten conjugate was used for immunization, followed by the hybridoma technique. Enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of LicoA was developed using a constructed antibody. The assay validation results were highly specific for the target compound, but minimally cross-reactive with the structure-related substances. After optimal conditions, the detection limit was 4.32 ng/mL and the quantification limit was 6.84-107.21 ng/mL. As a result, the developed ELISA was applied to determine the concentration of LicoA in raw licorice and marketed samples. This assay will aid in the quality control of licorice and derived products, as it can be used to identify plant species and determine bioactive markers.
•Heavy chain constant region 1 (CH1) influence heavy and light chains assembly.•Modified Fab (IgG1 subtype) improved in vitro refolding and intracellular folding.•Modified Fab (IgG1 subtype) improved ...reactivity.
Formation of disulfide bonds between heavy and light chains is challenge for production of recombinant immunoglobulin G (IgG) and fragment antigen-binding (Fab). Insufficient formation of the bond influences productive yield and quality of recombinant IgG and Fab. To investigate how amino acid sequences of the first heavy chain constant region (CH1) effect on assembly between heavy chain (VH-CH1 or Fd) and light chain (VL-CL), the CH1 gene sequence of an anti-paclitaxel fragment antigen-binding (anti-PT Fab IgG2a) was modified using the splicing by overlap extension-polymerase chain reaction (SOE-PCR) to be gene sequences encoding amino acid sequence of IgG1, IgG2b, and IgG3 subtypes. These anti-PT Fabs were expressed in Escherichia coli and silkworm larva expression systems. The efficiency of the assembly between VH-CH1 and VL-CL was evaluated. Based on sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis, assembly between heavy of IgG1 subtype and light chain was found to be superior over other subtypes using all tested expression systems. Furthermore, reactivity analysis using an enzyme-linked immunosorbent assay (ELISA) revealed that anti-PT Fab IgG1 subtype showed the highest reactivity against target compound paclitaxel. The folding efficiency and reactivity of the anti-PT Fab was influenced by CH1 amino acid sequence, which raises the possibility that this modification can be applied to improve recombinant Fab production.
Due to the highly specific binding between an antibody and its target, superior analytical performances was obtained by immunoassays for phytochemical analysis over conventional chromatographic ...techniques. Here, we describe a simple method for producing a functional single-chain variable fragment (scFv) antibody against ganoderic acid A (GAA), a pharmacologically active metabolite from Ganoderma lingzhi. The Escherichia coli BL21(DE3) strain produced a large amount of anti-GAA scFv. However, in vitro refolding steps, which partially recovered the reactivity of the scFv, were required. Interestingly, the functional scFv was expressed as a soluble and active form in the cytoplasm of an engineered E. coli SHuffle® strain. Purified anti-GAA scFv, which yielded 2.56 mg from 1 L of culture medium, was obtained from simple and inexpensive procedures for expression and purification. The anti-GAA scFv-based indirect competitive enzyme-linked immunosorbent assay (icELISA) exhibited high sensitivity (linearity: 0.078–1.25 µg/mL) with precision (CV: ≤6.20%) and reliability (recovery: 100.1–101.8%) for GAA determination. In summary, the approach described here is an inexpensive, simple, and efficient expression system that extends the application of anti-GAA scFv-based immunoassays. In addition, when in vitro refolding steps can be skipped, the cost and complexity of scFv antibody production can be minimized.
Pueraria candollei
is an ingredient of Thai herbal medicine, dietary supplements, and cosmetics. The in vitro and in vivo studies of this plant supported anti-osteoporotic activity and used for ...hormone replacement therapy. Deoxymiroestrol shows the most potent phytoconstituent in tuberous root of
P. candollei
with estrogenic activity
.
The quality controls are important for good agricultural practice (GAP) and good manufacturing practice (GMP) of plant-derived raw materials. The rapid detection of lateral flow immunoassay (LFIA) using colloidal gold is simply method, easy visualize detection and produce less waste than conventional chromatographic detection. In this study, LFIA for qualitative detection of deoxymiroestrol using antigen-binding fragment antibody (Fab) was developed. The result showed that the developed LFIA displays specific detection of deoxymiroestrol. Cross reactivity of this method was analyzed with miroestrol, isomiroestrol and methylisomiroestrol which showed 39.97%, 7.71% and 5.72%, respectively. After optimal condition, limit of detection (LOD) for deoxymiroestrol is 250 ng/ml. Plant samples were applied to strip test compare with indirect competitive ELISA using polyclonal antibody to confirm the application of LFIA. The results of LFIA method were comparable with those from ELISA. This developed lateral flow immunoassay can apply to detect deoxymiroestrol for the rapid testing. The developed method can use for quality control in plant samples as deoxymiroestrol is biomarker compound in
P. candollei
.
Graphic abstract
•One-step immunochromatographic assay (ICA) was developed for soy isoflavone glycosides.•Limit of detection of ICA system developed is ∼125ngmL−1.•Semi-quantitative analysis of DZ and GEN can be also ...performed by ICA developed.
Daidzin (DZ) and genistin (GEN) are two major soy isoflavone glycosides isolated from soybeans. Soy products containing isoflavones have recently been widely accepted for commercial use. However, the Japanese Government has suggested that soy isoflavone intake should be limited because of their estrogenic effects due to their interactions with estrogen receptors. In this study, we established a one-step indirect competitive immunochromatographic assay (ICA) for rapid and sensitive detection of total isoflavone glycosides (DZ and GEN) using gold nanoparticles conjugated with a monoclonal antibody against DZ. This assay was able to be completed in 15min following the immersion of a test strip in an analyte solution. Furthermore, the limit of detection for the total amount of isoflavone glycosides was ∼125ngmL−1. Considering that the major soy isoflavone glycosides found in soy products are DZ and GEN, this study demonstrates the potential use of ICA for the assessments of over consumption of isoflavones in soy supplements and foods, which would increase the safe dietary intake of soy products.
The peptide linker between variable domains of heavy (VH) and light (VL) chains is one of important factors that influence the characteristics of scFv, including binding activity and specificity ...against target antigen. The scFvs against daidzin (DZ-scFvs) with different linker lengths were constructed in the format of VH-(GGGGS)n-VL (n = 1, 3, 5, and 7). They were expressed in the hemolymph of silkworm larvae using the Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid DNA system, and their reactivity against daidzin and related compounds were evaluated using an indirect competitive enzyme-linked immunosorbent assay (icELISA), which is applicable for quantitative analysis of daidzin. The results showed that the reactivity of scFvs against daidzin was increased, whereas specificity slightly decreased when their peptide linker was lengthened. These results suggested that the linker length of DZ-scFvs contributes to its reactivity. In addition, the results emphasize that the linker length could control the reactivity of DZ-scFvs.
PDF
