Conjugation of Colloidal Gold to Proteins

Oliver, Constance

doi:10.1007/978-1-59745-324-0_39

Cited by 33 publications

(15 citation statements)

References 4 publications

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“…PcPHB1 was labeled with 10-nm-diameter gold nanoparticles (Sigma) following a previously reported method (28). Briefly, the pH of the colloidal gold was adjusted to at least 0.5 higher than the pI of PcPHB1 by using 0.1 N HCl.…”

Section: Methodsmentioning

confidence: 99%

Prohibitin Interacts with Envelope Proteins of White Spot Syndrome Virus and Prevents Infection in the Red Swamp Crayfish, Procambarus clarkii

Lan

Sun

et al. 2013

J Virol

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b Prohibitins (PHBs) are ubiquitously expressed conserved proteins in eukaryotes that are associated with apoptosis, cancer formation, aging, stress responses, cell proliferation, and immune regulation. However, the function of PHBs in crustacean immunity remains largely unknown. In the present study, we identified a PHB in Procambarus clarkii red swamp crayfish, which was designated PcPHB1. PcPHB1 was widely distributed in several tissues, and its expression was significantly upregulated by white spot syndrome virus (WSSV) challenge at the mRNA level and the protein level. These observations prompted us to investigate the role of PcPHB1 in the crayfish antiviral response. Recombinant PcPHB1 (rPcPHB1) significantly reduced the amount of WSSV in crayfish and the mortality of WSSV-infected crayfish. The quantity of WSSV in PcPHB1 knockdown crayfish was increased compared with that in the controls. The effects of RNA silencing were rescued by rPcPHB1 reinjection. We further confirmed the interaction of PcPHB1 with the WSSV envelope proteins VP28, VP26, and VP24 using pulldown and far-Western overlay assays. Finally, we observed that the colloidal gold-labeled PcPHB1 was located on the outer surface of the WSSV, which suggests that PcPHB1 specifically binds to the envelope proteins of WSSV. VP28, VP26, and VP24 are structural envelope proteins and are essential for attachment and entry into crayfish cells. Therefore, PcPHB1 exerts its anti-WSSV effect by binding to VP28, VP26, and VP24, preventing viral infection. This study is the first report on the antiviral function of PHB in the innate immune system of crustaceans.

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Section: Methodsmentioning

confidence: 99%

Prohibitin Interacts with Envelope Proteins of White Spot Syndrome Virus and Prevents Infection in the Red Swamp Crayfish, Procambarus clarkii

Lan

Sun

et al. 2013

J Virol

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“…As a general rule, it is considered that for maximizing the interaction of a protein with GNPs, conjugation should be conducted adjusting the pH at approximately 0.5 pH unit above the isoelectric point of the protein to be conjugated [36]. However, a fine-tuning of the system is recommended by determining the optimal pH value for conjugation for each specific antibody to each GNP preparation in order to maximize the sensitivity of the resulting assay [35,37].…”

Section: S-and -Dr-gnps Behavior During Conjugation With Antibodiesmentioning

confidence: 99%

Multicolor immunochromatographic strip test based on gold nanoparticles for the determination of aflatoxin B1 and fumonisins

et al. 2017

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Graphical abstractBlue desert rose-like gold nanoparticles (DR-GNPs) were synthesized, characterized and applied as label for the ImmunoChromatographic Strip Test (ICST) technique, in which red spherical GNPs (sGNPs) are usually employed. The combined use of the blue DR-GNP and red s-GNPs allowed developing of an intuitive multicolor ICST for the simultaneous detection of Aflatoxin B1 and AbstractDesert rose-like gold nanoparticles (DR-GNPs) that exhibited a blue color due to the plasmon resonance band at 620 nm were prepared. The blue DR-GNPs were obtained through a seeding growth approach and characterized by UV-vis spectroscopy, transmission electron microscopy and dynamic light scattering. The DR-GNPs had a hydrodynamic diameter of ~72 nm and were used as labels for the antibody in an immunochromatographic strip test (ICST). Although the particular shape of DR-GNPs and their higher surface area with respect to spherical GNPs, they demonstrated to be effective as antibody labels. A multicolor ICST for aflatoxin B1 and fumonisins was developed that employs both blue DR-and red spherical GNPs. The multicolor ICST allows for simultaneous rapid determination of the two mycotoxins in maize flour, with visual cut-off levels at 2 µg kg -1 for aflatoxin B1 and 1000 µg kg -1 for fumonisins.

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“…An emerging detection technique uses fluorescent microspheres [ 18 , 19 ] instead of colloidal gold particles [ 20 ]. Reaction bands can be observed with the naked eye following illumination with an ordinary UV lamp or special equipment.…”

Section: Introductionmentioning

confidence: 99%

Comparison of two different combined test strips with fluorescent microspheres or colored microspheres as tracers for rotavirus and adenovirus detection

Jiang

Shi

Lin

et al. 2018

Virol J

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BackgroundRotavirus (RV) and enteric adenovirus (AdV) mainly cause infantile infectious gastroenteritis. Several separate test methods for the detection of RV or AdV are currently available, but few tests are able to simultaneously detect both RV and AdV viruses, especially in primary medical institutions.MethodsThe present study was mainly designed to compare the performance of two combined test strips for the detection of RV and AdV: a rotavirus–adenovirus strip with fluorescent microspheres for tracers (FMT); and the CerTest rotavirus–adenovirus blister strip with colored microspheres for tracers (CMT). To test the strips cultures of RV, AdV and from other enteric pathogens were used, in addition to 350 stool specimens from 45 symptomatic patients with gastrointestinal infections.ResultsDetection thresholds for RV and AdV cultures using serial dilutions showed that the sensitivity of FMT was significantly higher than that of CMT (both P < 0.05). Specificity evaluation demonstrated that with culture mixtures of Coxsackie (A16), ECHO (type30), and entero- (EV71) viruses there was no detection of cross reaction using the two test strips, i.e., all the results were negative. With regard to the detection of RV in 350 clinical specimens, the total coincidence rate was 92.9%, the positive coincidence rate was 98.2%, and the negative coincidence rate was 90.8%. With regard to AdV detection, the total coincidence rate was 95.4%, the positive coincidence rate was 95.2%, and the negative coincidence rate was 95.5%.ConclusionsFMT performed better than CMT with regard to the combined detection of RV and AdV.

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Conjugation of Colloidal Gold to Proteins

Cited by 33 publications

References 4 publications

Prohibitin Interacts with Envelope Proteins of White Spot Syndrome Virus and Prevents Infection in the Red Swamp Crayfish, Procambarus clarkii

Prohibitin Interacts with Envelope Proteins of White Spot Syndrome Virus and Prevents Infection in the Red Swamp Crayfish, Procambarus clarkii

Multicolor immunochromatographic strip test based on gold nanoparticles for the determination of aflatoxin B1 and fumonisins

Comparison of two different combined test strips with fluorescent microspheres or colored microspheres as tracers for rotavirus and adenovirus detection

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