Membrane Immunoassay: Principle and Applications of Spin Membrane Immunoassay *

Hsia, J.C.; Tan, C. T.

doi:10.1111/j.1749-6632.1978.tb22019.x

Cited by 33 publications

(3 citation statements)

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“…Abolition of such spin-spin interactions would result in a sharp increase in resonance peak intensity due to narrowing of the line width. This effect has been previously used in a quantitative spin-membrane immunoassay (Hsia & Tan, 1978). In the present study, the reduction of nitroxide moieties on PNA in the process of TPH oxidation in the ischemic heart would reduce this proximity effect and result in a sharp increase in signal intensity.…”

Section: Discussionmentioning

confidence: 81%

Electron Paramagnetic Resonance Imaging of Rat Heart with Nitroxide and Polynitroxyl-Albumin

Kuppusamy¹,

Wang²,

Zweíer³

et al. 1996

Biochemistry

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Electron paramagnetic resonance (EPR) imaging utilizing stable nitroxyl radicals is a promising technique for measuring free radical distribution, metabolism, and tissue oxygenation in organs and tissues [Kuppusamy, P., Chzhan, M., Vij, K., Shteynbuk, M., Lefer, D. J., Giannella, E., & Zweier, J. L. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 3388-3392]. However, the technique has been limited by the rapid reduction of nitroxide in vivo to its hydroxylamine derivative, a diamagnetic, EPR-inactive species. In this report a novel, polynitroxylated derivative of human serum albumin is shown to be capable of reoxidizing the hydroxylamine back to nitroxide in vivo. Polynitroxyl-albumin (PNA) is shown to be effective in maintaining the signal intensity of the nitroxide 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL or TPL) in the ischemic isolated rat heart, allowing the acquisition of high-resolution three-dimensional (3D) EPR images of the heart throughout a prolonged 2.5 h period of global cardiac ischemia. In serial transverse sections of the 3D image, TPL intensity maps of the heart showed cardiac structure with submillimeter resolution. TPL intensities in coronary arteries and myocardium showed that nitroxide concentration decreases with increasing distance from large blood vessels. These results demonstrate that EPR imaging in vivo is possible using nitroxides in conjunction with PNA. In addition to its utility in the emerging technology of EPR imaging, the greatly prolonged half-life of TPL observed in the presence of PNA may facilitate the therapeutic application of nitroxides in a variety of disease processes.

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Section: Discussionmentioning

confidence: 81%

Electron Paramagnetic Resonance Imaging of Rat Heart with Nitroxide and Polynitroxyl-Albumin

Kuppusamy¹,

Wang²,

Zweíer³

et al. 1996

Biochemistry

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“…Antibodies were titrated easily by the agglutination of liposomes containing the antigen, or optically by recording increased light absorption as liposomes are agglutinated by antibodies (3,5) . A third method involves lysis of liposomes by complement and specific antibodies using markers (36,60,61) to measure the degree of lysis . In our assay a colored substance, sodium dichromate, was used as a marker, and the assay was carried out in microequilibrium dialysis cells to minimize the quantities of liposomes, antibody, and complement needed .…”

Section: Discussionmentioning

confidence: 99%

Immunochemical studies of conjugates of isomaltosyl oligosaccharides to lipid. I. Antigenicity of the glycolipids and the production of specific antibodies in rabbits.

Wood¹,

Kabat

1981

The Journal of Experimental Medicine

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The antigenicity of lipids is still ill-defined, and there is very little evidence that any pure lipid by itself will act as an antigen . To produce an immune response they must be complexed with or coupled to carriers (1-6) . Only glycosphingolipids and phosphatides have been found to function as haptens when mixed with carriers such as serum albumin (7), methylated bovine serum albumin (8), or other auxillary lipids (9-11), or when the glycolipid was incorporated into liposomes (12) . Antibodies to glycolipids are usually directed primarily against the carbohydrate moiety (1, 7, 13, 14), and there is little evidence that antibodies can be formed to the free lipids even though free fatty acids were reported to cause inflammatory delayed-type allergic skin reactions (15, 16) .The purpose of this investigation was to synthesize and study the antigenicity and immunological properties of a series of glycolipid antigens prepared by coupling isomaltose oligosaccharides-varying in size from isomaltose to isomaltoheptaose-to a lipid carrier, stearylamine, and to compare the antigenicities of the different stearylisomaltosyl oligosaccharides thus obtained, either when injected alone or when incorporated into liposomes. Isomaltose oligosaccharides were chosen because they are the predominant structural units of a1--),6 dextran (17). Antibodies raised to them should cross-react with dextrans, and the heterogeneity of the response and the sizes of their antibody combining sites could be studied by quantitative precipitin and precipitin inhibition assays and could be compared with antidextran (18-22) .The stearyl-isomaltosyl oligosaccharides (stearyl-IM3 to IM7) when incorporated into liposomes were agglutinated in the presence of specific antibodies and were lysed ifcomplement was also present . When injected into rabbits they were antigenic, either emulsified directly with complete Freund's adjuvant (CFA) l or incorporated into * Aided by grants from the National Science Foundation (PCM 76-81029) and Cancer Center Support grant CA 13696 to the Cancer Center, Columbia University .f Portions of this work are from part I of a dissertation submitted by C . Wood in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences, Columbia University, New York .'Abbreviations used in this paper: BSA, bovine serum albumin ; CFA, complete Freund's adjuvant ; DNPCap-PE, DNP-e-aminocaproylphosphatidyl-ethanolamine ; yglo, gammaglobulin; IM2, isomaltose (DGlcal~6DGlc) ; IM3, isomaltotriose ; IM4, isomaltotetraose ; IM5, isomaltopentaose ; IM6, isomaltohexaose ; IM7, isomaltoheptaose ; PBS, phosphate-buffered saline (0 .01 M phosphate, 0 .8 percent sodium chloride, pH 7 .2) ; SDS, sodium dodecyl sulfate; SRBC, sheep erythrocytes ; TCA, trichloroacetic acid . 432J. Exp . MED .

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“…The main difficulty in the analysis of the antigen-antibody reaction is that lipids, unlike most proteins, sugars, or artificial antigens, are insoluble in aqueous solutions which are optimal for antibody activity and tend to form macromolecular aggregates of micellar nature. However, it has been shown that lipid antigens may be solubilized in a lipid bilayer membrane matrix to form sensitized liposomes (1)(2)(3)(4). The liposomes are artificial vesicles which consist of concentric shells of lipid bilayer, whose aqueous interspaces can contain trapped markers.…”

mentioning

confidence: 99%

Thin-layer potentiometric analysis of lipid antigen-antibody reaction by tetrapentylammonium (TPA+) ion loaded liposomes and TPA+ ion selective electrode

Shiba¹,

Umezawa²,

Watanabe³

et al. 1980

Anal. Chem.

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Membrane Immunoassay: Principle and Applications of Spin Membrane Immunoassay *

Cited by 33 publications

References 1 publication

Electron Paramagnetic Resonance Imaging of Rat Heart with Nitroxide and Polynitroxyl-Albumin

Electron Paramagnetic Resonance Imaging of Rat Heart with Nitroxide and Polynitroxyl-Albumin

Immunochemical studies of conjugates of isomaltosyl oligosaccharides to lipid. I. Antigenicity of the glycolipids and the production of specific antibodies in rabbits.

Thin-layer potentiometric analysis of lipid antigen-antibody reaction by tetrapentylammonium (TPA+) ion loaded liposomes and TPA+ ion selective electrode

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