The adsorptive characteristics of proteins for polystyrene and their significance in solid-phase immunoassays

Cantarero, Lara; Butler, John E.; Osborne, James C.

doi:10.1016/0003-2697(80)90473-x

Cited by 298 publications

(107 citation statements)

References 11 publications

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“…Using the bicinchoninic acid protein assay, we estimated the amount of C225 attached to the HAuNS to be 124 antibodies/nanoshell. The amount of antibody that would form a monolayer on the surface of HAuNS could be calculated using saturation capacity data of f2.5 mg/m 2 for IgG (or f100 nm 2 /antibody) provided by Cantarero et al (32). Thus, based on the spherical shape and radius of 15 nm, the number of antibodies on the gold surface was 28 antibodies/nanoshell calculated by dividing the total surface area of a nanoshell by the head group surface area of an antibody.…”

Section: Discussionmentioning

confidence: 99%

In vitro and in vivo targeting of hollow gold nanoshells directed at epidermal growth factor receptor for photothermal ablation therapy

Melancon¹,

Lü²,

Liu³

et al. 2008

Molecular Cancer Therapeutics

380

422

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Section: Discussionmentioning

confidence: 99%

In vitro and in vivo targeting of hollow gold nanoshells directed at epidermal growth factor receptor for photothermal ablation therapy

Melancon¹,

Lü²,

Liu³

et al. 2008

Molecular Cancer Therapeutics

380

422

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“…This amount is usually estimated and depends upon factors such as the molecular weight of the protein, its relative affinity for the particle and the particle size [43]. While the amount of protein required to achieve surface saturation can be calculated, the actual protein concentration in the reaction may be substantially higher than the "monolayer = 1 equivalent of protein" value.…”

Section: Concentration Of the Biomoleculementioning

confidence: 99%

A comparison of mono and multivalent linkers and their effect on the colloidal stability of nanoparticle and immunoassays performance

Gubala

Guével

Nooney

et al. 2010

Talanta

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When designing devices for biomedical diagnostics, increasing the signal to noise ratio is often critical for achieving clinically relevant sensitivity and limits of detection (LOD). In antibodybased assays, the measured signal can be amplified through the replacement of molecular fluorophores with doped nanoparticles (NP). However, the benefits of using NPs can only be realized if the NPs are coated efficiently with detection antibody, have good colloidal stability and the ratio of specific to non-specific binding (NSB) is high enough. The main focus of this paper is on the optimization of the bioconjugation protocol for antibody labeling of NPs leading to improved assay performance. Two types of linkers were used: monovalent linkers (glutaraldehyde; sulfo-SMCC; and sulfo-SIAB), and three generations of dendrimers endowed with multivalent carboxylic functionality. Overall, the NP-IgG conjugates prepared using multivalent linkers showed a significantly lower LOD and higher sensitivity than their homo-or hetero-functional counterparts. The multivalent dendrimers also improved NP stability and reduced aggregation. Moreover, the dendrimers showed a higher reactivity with biological material, a feature that could significantly reduce the cost of high-throughput biodiagnostics tests. Introduction:

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“…Generation and purification of the recombinant rhodostomin proteins were done according to a previous report (18). All beads used in this study were polystyrene beads (Bangs Laboratories); the coating with recombinant rhodostomin proteins was done by the hydrophobic adsorption method (19). Essentially, beads were incubated with 10 g/ml proteins in phosphate-buffered saline at 4°C overnight with gentle mixing, washed with phosphate-buffered saline for three times, and further blocked with 5% heat-treated bovine serum albumin at 4°C for 2 h.…”

Section: Methodsmentioning

confidence: 99%

Stepped Changes of Monovalent Ligand-binding Force during Ligand-induced Clustering of Integrin αIIBβ3

Hsieh¹,

Chang

Pai

et al. 2006

Journal of Biological Chemistry

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Recent evidence demonstrated that conformational changes of the integrin during receptor activation affected its binding to extracellular matrix; however, experimental assessment of ligand-receptor binding following the initial molecular interaction has rarely been carried out at a single-molecule resolution. In the present study, laser tweezers were used to measure the binding force exerted by a live Chinese hamster ovary cell that expressed integrin ␣ IIb ␤ 3 (CHO ␣ IIb ␤ 3 ), to the bead carrier coated with the snake venom rhodostomin that served as an activated ligand for integrin ␣ IIb ␤ 3 . A progressive increase of total binding force over time was noticed when the bead interacted with the CHO ␣ IIb ␤ 3 cell; such an increase was due mainly to the recruitment of more integrin molecules to the bead-cell interface. When the binding strength exerted by a single ligandreceptor pair was derived from the "polyvalent" measurements, surprisingly, a stepped decrease of the "monovalent binding force" was noted (from 4.15 to 2.54 piconewtons (pN)); such decrease appeared to occur during the ligand-induced integrin clustering process. On the other hand, the mutant rhodostomin defective in clustering integrins exhibited only one (1.81 pN) unit binding strength.Since the early recognition of integrin as an integral membrane complex involved in the trans-membrane association between the extracellular matrix and the cell, integrin heterodimers have been found to play important roles in controlling various steps that regulate processes as diverse as proliferation, development and differentiation, cell migration, and carcinogenesis (1). The initiation of the integrin-mediated activities involves at least two steps, namely, receptor activation and clustering. Taking platelet cells as an example, the integrin ␣ IIb ␤ 3 proteins on the membrane of a resting platelet bind only loosely to fibrinogen (FBN).3 Stimulation (or activation) of the platelets with agonists (such as ADP or thrombin) induces an inside-out signaling process that confers on integrin ␣ IIb ␤ 3 a stronger binding to FBN, resulting in the onset of platelet aggregation. Subsequent integrin clustering triggers complex intracellular signaling pathways that regulate the extent of the irreversible platelet aggregation and clot retraction (2). Using advanced structural biology techniques, several recent reports have provided convincing evidence that indicates the adoption of different molecular conformations by integrin heterodimers when the inside-out signaling pathway is invoked (3-5). Such structural changes are thought to be the mechanisms underlying increased ligand binding by an integrin during the activation process. Whether or not ligand binding affinity is further modulated during the subsequent integrin clustering process has remained largely elusive (6).To focus this research on the ligand-receptor interaction during the receptor clustering, we utilized snake venom rhodostomin from Agkistrodon rhodostoma as the ligand for integrin ␣ IIb ␤ 3 . Rhodostomin...

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The adsorptive characteristics of proteins for polystyrene and their significance in solid-phase immunoassays

Cited by 298 publications

References 11 publications

In vitro and in vivo targeting of hollow gold nanoshells directed at epidermal growth factor receptor for photothermal ablation therapy

In vitro and in vivo targeting of hollow gold nanoshells directed at epidermal growth factor receptor for photothermal ablation therapy

A comparison of mono and multivalent linkers and their effect on the colloidal stability of nanoparticle and immunoassays performance

Stepped Changes of Monovalent Ligand-binding Force during Ligand-induced Clustering of Integrin αIIBβ3

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