Gold Nanoparticle Based Immunostrip Assay Method for Detection of Protein-A

2012 ◽  
Vol 229-231 ◽  
pp. 260-266 ◽  
Author(s):  
Ajit Zambre ◽  
Nripen Chanda ◽  
Sudhirdas Prayaga ◽  
Rosana Almudhafar ◽  
Raghuraman Kannan ◽  
...  

We have successfully developed gold nanoparticle based immunostrip assay to detect protein-A (PA). Rabbit polyclonal antibody IGg (αPA) that has affinity to PA was conjugated to gold nanoparticles (GNPs) and the gold nanoconjugate (αPA-GNP) was used to detect protein-A by simple immunostrip assay method. ELISA experiments were used to confirm the retention of binding affinity of antibody towards protein-A after conjugation with gold nanoparticles.

Nanoscale ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 3665-3673 ◽  
Author(s):  
Hanane Moustaoui ◽  
Justine Saber ◽  
Ines Djeddi ◽  
Qiqian Liu ◽  
Dania Movia ◽  
...  

Gold nanoparticle interaction with proteins is characterized by using scattering correlation spectroscopy. Protein orientation and binding affinity regarding the nanoparticle surface are discussed.


2020 ◽  
Vol 16 (2) ◽  
pp. 204-213 ◽  
Author(s):  
Melissa A. Vetten ◽  
Mary Gulumian

Background: Endotoxin-free engineered nanoparticle suspensions are imperative for their successful applications in the field of nanomedicine as well as in the investigations in their toxicity. Gold nanoparticles are known to interfere with various in vitro assays due to their optical properties and potential for surface reactivity. In vitro endotoxin testing assays are known to be susceptible to interference caused by the sample being tested. Objective: This study aimed to identify a preferred assay for the testing of endotoxin contamination in gold nanoparticle suspensions. Methods: The interference by gold nanoparticles on three assays namely, the commonly used limulus amebocyte lysate chromogenic assay, the limulus amebocyte lysate gel-clot method, and the less common recombinant Factor C (rFC) assay, was tested. Results: Possible interference could be observed with all three assays. The interference with the absorbance- based chromogenic assay could not be overcome by dilution; whilst the qualitative nature of the gel-clot assay excluded the possibility of distinguishing between a false positive result due to enhancement of the sensitivity of the assay, and genuine endotoxin contamination. However, interference with the rFC assay was easily overcome through dilution. Conclusion: The rFC assay is recommended as an option for endotoxin contamination detection in gold nanoparticle suspensions.


2021 ◽  
Author(s):  
Yiren Cao ◽  
Jinjun Wu ◽  
Bo Pang ◽  
Hongquan Zhang ◽  
X. Chris Le

The trans-cleavage activity of the target-activated CRISPR-Cas12a liberated an RNA crosslinker from a molecular transducer, which facilitated assembly of gold nanoparticles. Integration of the molecular transducer with isothermal amplification and...


2021 ◽  
Author(s):  
Min Chen ◽  
Leiqing Pan ◽  
K. Tu

A simple and quick responsive fluorescent biosensor for Salmonella typhimurium detection based on the recognition of aptamer coupled with alendronic acid (ADA)@upconversion nanoparticles (UCNPs) and gold nanoparticle (AuNPs) has been...


Metals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 472
Author(s):  
Qunying Yuan ◽  
Manjula Bomma ◽  
Zhigang Xiao

Phytochelatins, the enzymatic products of phytochelatin synthase, play a principal role in protecting the plants from heavy metal and metalloid toxicity due to their ability to scavenge metal ions. In the present study, we investigated the capacity of soluble intracellular extracts from E. coli cells expressing R. tropici phytochelatin synthase to synthesize gold nanoparticle. We discovered that the reaction mediated by soluble extracts from the recombinant E. coli cells had a higher yield of gold nanoparticles, compared to that from the control cells. The compositional and morphological properties of the gold nanoparticles synthesized by the intracellular extracts from recombinant cells and control cells were similar. In addition, this extracellular nanoparticle synthesis method produced purer gold nanoparticles, avoiding the isolation of nanoparticles from cellular debris when whole cells are used to synthesize nanoparticles. Our results suggested that phytochelatins can improve the efficiency of gold nanoparticle synthesis mediated by bacterial soluble intracellular extracts, and the potential of extracellular nanoparticle synthesis platform for the production of nanoparticles in large quantity and pure form is worth further investigation.


The Analyst ◽  
2017 ◽  
Vol 142 (17) ◽  
pp. 3261-3271 ◽  
Author(s):  
John A. Hondred ◽  
Joyce C. Breger ◽  
Nate T. Garland ◽  
Eunkeu Oh ◽  
Kimihiro Susumu ◽  
...  

A significant enzymatic enhancement of phosphotriesterase has been demonstrated when immobilized on various sized gold nanoparticles.


2010 ◽  
Vol 115 (3) ◽  
pp. 653-660 ◽  
Author(s):  
Siyam M. Ansar ◽  
Rukshani Haputhanthri ◽  
Bradley Edmonds ◽  
Dong Liu ◽  
Leyuan Yu ◽  
...  

2016 ◽  
Vol 18 (3) ◽  
pp. 1953-1960 ◽  
Author(s):  
M. C. R. González ◽  
A. G. Orive ◽  
R. C. Salvarezza ◽  
A. H. Creus

Gold nanoparticle electrodeposition on a modified HOPG surface with a monolayer organic film based on aryl diazonium chemistry has been studied.


1995 ◽  
Vol 309 (1) ◽  
pp. 141-144 ◽  
Author(s):  
F W Holtsberg ◽  
L E Ozgur ◽  
D E Garsetti ◽  
J Myers ◽  
R W Egan ◽  
...  

The supernatant fraction from lysed human eosinophils, when separated by gel-filtration chromatography, contains a protein with lysophospholipase activity of approximate molecular mass 74 kDa. This mass differs substantially from the 17 kDa of a previously cloned eosinophil lysophospholipase (Charcot-Leyden crystal protein), but is similar to that reported for a pancreatic enzyme. We have therefore further characterized this pancreatic-like lysophospholipase in human eosinophils. A rabbit polyclonal antibody was produced against a synthetic peptide consisting of amino acids 325-349 from the 74 kDa rat pancreatic lysophospholipase. Western-blot analysis of eosinophil extracts indicate that this antibody recognizes a single 74 kDa band in these preparations. Incubation of the supernatant fraction from sonified eosinophils with this antibody, followed by precipitation of antibody-antigen complexes with Protein A, removes the majority of the lysophospholipase activity. Indirect immunofluorescence examination with this antibody indicates this protein to be localized to granules of eosinophils and not in other leucocytes. Moreover, reverse transcriptase PCR of polyadenylated RNA from eosinophils and from rat pancreatic tissue with primers to rat pancreatic lysophospholipase resulted in readily detectable 1 kb DNA products in both samples. Sequencing revealed this DNA fragment to be identical with the human pancreatic lysophospholipase cDNA sequence. Taken together, these data indicate that eosinophils contain a lysophospholipase that is similar to the human pancreatic enzyme.


Nanoscale ◽  
2015 ◽  
Vol 7 (26) ◽  
pp. 11420-11432 ◽  
Author(s):  
Prabhani U. Atukorale ◽  
Yu-Sang Yang ◽  
Ahmet Bekdemir ◽  
Randy P. Carney ◽  
Paulo J. Silva ◽  
...  

Amphiphilic gold nanoparticles spontaneously insert into erythrocyte membranes; we characterize this association as a function of key plasma membrane components.


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