Low-Level Detection of Poly(amidoamine) PAMAM Dendrimers Using Immunoimaging Scanning Probe Microscopy

Cason, Chevelle A.; Fabré, Thomas A.; Buhrlage, Andrew; Haik, Kristi L.; Bullen, Heather A.

doi:10.1155/2012/341260

Cited by 8 publications

(6 citation statements)

References 42 publications

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“…As a result, the presence of dense concentration and globular shape of dendrimer might have provided better interaction between antigen and sensing layer 79 . Therefore, 10 mM PAMAM dendrimer was established as optimal concentration, which is in agreement with those reported in the literature 80 .…”

Section: Resultssupporting

confidence: 89%

Sensitive Detection of Dengue Virus Type 2 E-Proteins Signals Using Self-Assembled Monolayers/Reduced Graphene Oxide-PAMAM Dendrimer Thin Film-SPR Optical Sensor

Omar

Fen

Abdullah

et al. 2020

Sci Rep

119

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In this work, sensitive detection of dengue virus type 2 E-proteins (DENV-2 E-proteins) was performed in the range of 0.08 pM to 0.5 pM. The successful DENV detection at very low concentration is a matter of concern for targeting the early detection after the onset of dengue symptoms. Here, we developed a SPR sensor based on self-assembled monolayer/reduced graphene oxide-polyamidoamine dendrimer (SAM/NH 2 rGO/PAMAM) thin film to detect DENV-2 E-proteins. Surface characterizations involving X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR) confirms the incorporation of NH 2 rGO-PAMAM nanoparticles in the prepared sensor films. The specificity, sensitivity, binding affinity, and selectivity of the SPR sensor were then evaluated. Results indicated that the variation of the sensing layer due to different spin speed, time incubation, and concentration provided a better interaction between the analyte and sensing layer. The linear dependence of the SPR sensor showed good linearity (R 2 = 0.92) with the lowest detection of 0.08 pM DENV-2 E-proteins. By using the Langmuir model, the equilibrium association constant was obtained at very high value of 6.6844 TM −1 (R 2 = 0.99). High selectivity of the SPR sensor towards DENV-2 E-proteins was achieved in the presence of other competitors. Dengue virus (DENV) is the most common arthropod-borne viral disease that poses a serious global problem. According to World Health Organization (WHO), the dengue virus is the leading cause of death of 22 000, annually. As of today, the need for hospitalization and medical treatment are constantly dense due to the fact that 390 million people in the world are still infected with DENV 1. Its four distinct serotypes (DENV-1. DENV-2, DENV-3, and DENV-4) are capable of causing a range of clinical symptoms ranging from mild fevers to the severe dengue haemorrhagic fever (DHF) and be potentially life-threatening 2-10. Despite its large burden to human health, no effective vaccine and antiviral therapy are available for the virus 11,12. Early treatment for DENV is only by maintaining the body fluid of the patient, as it is critical in fighting the severe symptoms of DENV 13,14. Hence, an early, rapid, and accurate diagnosis at the onset of infection is the demand of the day in the most epidemic settings. Present discoveries in dengue diagnostics that can help in the early diagnosis are targeting the host-virus itself. DENV consists of a single-stranded positive-sense RNA virus that encodes 10 different types of proteins. Seven of

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Section: Resultssupporting

confidence: 89%

Sensitive Detection of Dengue Virus Type 2 E-Proteins Signals Using Self-Assembled Monolayers/Reduced Graphene Oxide-PAMAM Dendrimer Thin Film-SPR Optical Sensor

Omar

Fen

Abdullah

et al. 2020

Sci Rep

119

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“…Cross-sectional line scans along the dotted segments in Figure 4b,c are illustrated in Figure 4d. The particle heights obtained from four distinct particles represented in Figure 4d range from 2.7–10 nm which agrees with dendrimer size obtained by AFM analysis reported by Li et al and with dendrimer analysis observed in our previous work [38,68]. Feature width, from Figure 4d, is 30–50 nm which is 7–13 times larger than the expected diameter of G4 PAMAM dendrimers, 4 nm [69].…”

Section: Resultssupporting

confidence: 89%

Analysis of Biotinylated Generation 4 Poly(amidoamine) (PAMAM) Dendrimer Distribution in the Rat Brain and Toxicity in a Cellular Model of the Blood-Brain Barrier

et al. 2013

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Dendrimers are highly customizable nanopolymers with qualities that make them ideal for drug delivery. The high binding affinity of biotin/avidin provides a useful approach to fluorescently label synthesized dendrimer-conjugates in cells and tissues. In addition, biotin may facilitate delivery of dendrimers through the blood-brain barrier (BBB) via carrier-mediated endocytosis. The purpose of this research was to: (1) measure toxicity using lactate dehydrogenase (LDH) assays of generation (G)4 biotinylated and non-biotinylated poly(amidoamine) (PAMAM) dendrimers in a co-culture model of the BBB, (2) determine distribution of dendrimers in the rat brain, kidney, and liver following systemic administration of dendrimers, and (3) conduct atomic force microscopy (AFM) on rat brain sections following systemic administration of dendrimers. LDH measurements showed that biotinylated dendrimers were toxic to cell co-culture after 48 h of treatment. Distribution studies showed evidence of biotinylated and non-biotinylated PAMAM OPEN ACCESSMolecules 2013, 18 11538 dendrimers in brain. AFM studies showed evidence of dendrimers only in brain tissue of treated rats. These results indicate that biotinylation does not decrease toxicity associated with PAMAM dendrimers and that biotinylated PAMAM dendrimers distribute in the brain. Furthermore, this article provides evidence of nanoparticles in brain tissue following systemic administration of nanoparticles supported by both fluorescence microscopy and AFM.

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“…Most existing instrumental methods, e.g., IR spectroscopy, NMR spectroscopy, size-exclusion chromatography, and mass spectrometry (MS) especially with matrix-assisted laser desorption/ionization, have been applied in investigations of the synthesis of dendrimers to optimize aspects such as control of structural defects and functionalization of the surface [2,3,[8][9][10][11][12][13][14]. Sophisticated microscopy (image-based) techniques or chromatographic systems are being used as the tool of choice for detection, quantification, and localization of nanoparticles in cells to describe their metabolic pathways [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

“…Sophisticated microscopy (image-based) techniques or chromatographic systems are being used as the tool of choice for detection, quantification, and localization of nanoparticles in cells to describe their metabolic pathways [8][9][10]. Soft ionization technique such as electrospray ionization (ESI) MS have been shown to efficiently promote ionization of dendrimers ranging from low to high generations.…”

Section: Introductionmentioning

confidence: 99%

In vitro dose–response effects of poly(amidoamine) dendrimers [amino-terminated and surface-modified with N-(2-hydroxydodecyl) groups] and quantitative determination by a liquid chromatography–hybrid quadrupole/time-of-flight mass spectrometry based method

et al. 2012

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This article presents a dose-response study of the effects of two types of third-generation (G3) and fourth-generation poly(amidoamine) (PAMAM) dendrimers on two cell lines (RTG-2 and H4IIE) by in vitro cytotoxicity assays with 3-(4,5-dimethylthizol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), neutral red uptake (NRU), and lactate dehydrogenase (LDH) assays. We particularly investigated the potential cytotoxic effect of positive surface charge, which a cationic amino-terminated PAMAM dendrimer can display, on the marked ability of PAMAM dendrimers to cross the cell membrane compared with PAMAM dendrimers functionalized with chains of N-(2-hydroxydodecyl). Quantification of dose-response effects was performed by use of mass spectrometry analysis. The analytical method using liquid chromatography-hybrid quadrupole/time-of-flight mass spectrometry that we developed allowed characterization of defective dendrimers instead of "ideal structures." Identification was based on accurate mass measurement, assignment of elemental composition, and the fully resolved (13)C/(12)C isotopic clusters of the multiply charged ions of PAMAM dendrimers. Validation of the liquid chromatography-mass spectrometry method made possible reliable and reproducible quantification of the extracellular and intracellular concentration of dendrimers at a micromolar level (limits of detection from 0.14 to 1.34 μM and from 0.43 to 1.82 μM in standard and culture medium, respectively). A higher cytotoxicity was found with the H4IIE cell line for surface-modified PAMAM dendrimers. The LDH assay was significantly more sensitive than the MTT and NRU assays, with half-maximal inhibitory concentrations (IC(50)) of 12.96 and 38.31 μg mL(-1) for surface-modified G3 and G4 dendrimers, respectively. No cytotoxic effects, in terms of IC(50), of amino-terminated PAMAM dendrimers were observed on both H4IIE and RTG-2 cells when the concentration was below 500 μg mL(-1) for G3 and G4 dendrimers.

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Low-Level Detection of Poly(amidoamine) PAMAM Dendrimers Using Immunoimaging Scanning Probe Microscopy

Cited by 8 publications

References 42 publications

Sensitive Detection of Dengue Virus Type 2 E-Proteins Signals Using Self-Assembled Monolayers/Reduced Graphene Oxide-PAMAM Dendrimer Thin Film-SPR Optical Sensor

Sensitive Detection of Dengue Virus Type 2 E-Proteins Signals Using Self-Assembled Monolayers/Reduced Graphene Oxide-PAMAM Dendrimer Thin Film-SPR Optical Sensor

Analysis of Biotinylated Generation 4 Poly(amidoamine) (PAMAM) Dendrimer Distribution in the Rat Brain and Toxicity in a Cellular Model of the Blood-Brain Barrier

In vitro dose–response effects of poly(amidoamine) dendrimers [amino-terminated and surface-modified with N-(2-hydroxydodecyl) groups] and quantitative determination by a liquid chromatography–hybrid quadrupole/time-of-flight mass spectrometry based method

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