Use of microwell plates carrying hydrazide groups to enhance antibody immobilization in enzyme immunoassays

Brillhart, Kurt L.; Ngo, T. T.

doi:10.1016/0022-1759(91)90225-5

Cited by 27 publications

(13 citation statements)

References 13 publications

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“…Immobilization of Abs onto plate microwells with hydrazide surface (HZ wells) (carbohydrate-binding 8-well Strip Plate, catalog no. 2508; Costar), which allows site-specific binding of Abs through the carbohydrate moieties of the Fc region (35), was performed exactly as described by Menegazzi et al (9). Supernatant fluids or cell lysates were diluted 300-fold in PBS containing 0.1% (w/v) BSA and 0.1% (v/v) Tween 20 (Merck) (PBS-T-BSA).…”

Section: Assay Of Lactoferrin Releasementioning

confidence: 99%

Chloride Movements in Human Neutrophils during Phagocytosis: Characterization and Relationship to Granule Release

Busetto

Trevisan

Decleva

et al. 2007

The Journal of Immunology

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Chloride ion efflux is an early event occurring after exposure of human neutrophils to several soluble agonists. Under these circumstances, a rapid and reversible fall in the high basal intracellular chloride (Cl−i) levels is observed. This event is thought to play a crucial role in the modulation of several critical neutrophil responses including activation and up-regulation of adhesion molecules, cell attachment and spreading, cytoplasmic alkalinization, and activation of the respiratory burst. At present, however, no data are available on chloride ion movements during neutrophil phagocytosis. In this study, we provide evidence that phagocytosis of Candida albicans opsonized with either whole serum, complement-derived opsonins, or purified human IgG elicits an early and long-lasting Cl− efflux accompanied by a marked, irreversible loss of Cl−i. Simultaneous assessment of Cl− efflux and phagocytosis in cytochalasin D-treated neutrophils indicated that Cl− efflux occurs without particle ingestion. These results suggest that engagement of immune receptors is sufficient to promote chloride ion movements. Several structurally unrelated chloride channel blockers inhibited phagocytosis-induced Cl− efflux as well as the release of azurophilic—but not specific—granules. It implicates that different neutrophil secretory compartments display distinct sensitivity to Cl−i modifications. Intriguingly, inhibitors of Cl− exchange inhibited cytosolic Ca2+ elevation, whereas Cl− efflux was not impaired in Ca2+-depleted neutrophils. We also show that FcγR(s)- and CR3/CR1-mediated Cl− efflux appears to be dependent on protein tyrosine phosphorylation but independent of PI3K and phospholipase C activation.

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Section: Assay Of Lactoferrin Releasementioning

confidence: 99%

Chloride Movements in Human Neutrophils during Phagocytosis: Characterization and Relationship to Granule Release

Busetto

Trevisan

Decleva

et al. 2007

The Journal of Immunology

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“…16–21 Selective binding to the Fc region of an Ab is beneficial for maximizing the biological activity of the Fab binding regions. 19,20,22–24 This conjugation strategy allows for the multiplexing of various glycosylated Abs on a single nanoparticle that can be used for delivery and monitoring of therapeutic agents in vivo. 19,20,25,26 Thus, gold surface can serve as a high versatile substrate for binding of multiple types of functional biomolecules with relatively simple conjugation chemistry.…”

Section: Introductionmentioning

confidence: 99%

Selective Targeting of Antibody Conjugated Multifunctional Nanoclusters (Nanoroses) to Epidermal Growth Factor Receptors in Cancer Cells

Tam

Willsey

et al. 2011

Langmuir

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The ability of smaller than 100 nm antibody (Ab) nanoparticle conjugates to target and modulate the biology of specific cell types may enable major advancements in cellular imaging and therapy in cancer. A key challenge is to load a high degree of targeting, imaging, and therapeutic functionality into small, yet stable particles. A versatile method called thin autocatalytic growth on substrate (TAGs) has been developed in our previous study to form ultra-thin and asymmetric gold coatings on iron oxide nanocluster cores producing exceptional near infrared (NIR) absorbance. AlexaFluor 488 labeled Abs were used to correlate the number of Abs conjugated to iron oxide/gold nanoclusters (nanoroses) with the hydrodynamic size. A transition from sub-monolayer to multilayer aggregates of Abs on the nanorose surface was observed for 54 Abs and an overall particle diameter of ~60 to 65 nm. The hydrodynamic diameter indicated coverage of a monolayer of 54 Abs, in agreement with the prediction of a geometric model, by assuming a circular footprint of 16.9 nm diameter per Ab molecule. The targeting efficacy of nanoclusters conjugated with monoclonal Abs specific for epidermal growth factor receptor (EGFR) was evaluated in A431 cancer cells using dark field microscopy and atomic absorbance spectrometry (AAS) analysis. Intense NIR scattering was achieved from both high uptake of nanoclusters in cells and high intrinsic NIR absorbance of individual nanoclusters. Dual mode imaging with dark field reflectance microscopy and fluorescence microscopy indicates the Abs remained attached to the Au surfaces upon the uptake by the cancer cells. The ability to load intense multifunctionality, specifically strong NIR absorbance, conjugation of an Ab monolayer in addition to a strong r2 MRI contrast that was previously demonstrated in a total particle size of only 63 nm, is an important step forward in development of theranostic agents for combined molecular specific imaging and therapy.

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“…Although bio-rabbit anti-mouse (34 mg/ml) and rabbit anti-sheep IgG molecules effectively bind to solid surfaces (31), they may (12 mg/ml) were obtained from Sigma. Goat anti-mouse denature in the process, thereby losing their biospecific activ-antibodies (molecular weight 150,000 Da) specific for the ity (32). Covalent binding of biomolecules to the surface is mouse F (ab)2 and F (c) IgG regions were supplied by Sigma favored and procedures resulting in minimal loss of activity as powders and made up to solutions of 2.3 mg/ml with have been employed (2, 5, 11,14,33,34).…”

Section: Introductionmentioning

confidence: 99%

Orientational Aspects of Antibody Immobilization and Immunological Activity on Quartz Crystal Microbalance Electrodes

Caruso

Rodda

Furlong

1996

Journal of Colloid and Interface Science

133

112

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Use of microwell plates carrying hydrazide groups to enhance antibody immobilization in enzyme immunoassays

Cited by 27 publications

References 13 publications

Chloride Movements in Human Neutrophils during Phagocytosis: Characterization and Relationship to Granule Release

Chloride Movements in Human Neutrophils during Phagocytosis: Characterization and Relationship to Granule Release

Selective Targeting of Antibody Conjugated Multifunctional Nanoclusters (Nanoroses) to Epidermal Growth Factor Receptors in Cancer Cells

Orientational Aspects of Antibody Immobilization and Immunological Activity on Quartz Crystal Microbalance Electrodes

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