Enzyme Immunoassay in Rapid Progress

Kato, Kanefusa; Hamaguchi, Yukio; Okawa, Sho; Ishikawa, Eiji; Kobayashi, Keizo; Katunuma, Nobuhiko

doi:10.1016/s0140-6736(77)91673-7

Cited by 26 publications

(8 citation statements)

References 3 publications

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“…The sandwich-type enzyme immunoassay system was composed of polystyrene balls (3.18 mm in diameter; Precision Plastic Ball Co., Chicago, IL) with immobilized antibody F(ab')2 fragments, and the same antibody Fab' fragments were labeled with j3-D-galactosidase from Escherichia coli. Details of immobilization of antibodies on polystyrene balls (Kato et al, 1977) and labeling of antibodies with /3-D-galactosidase (Kato et al, 1976) are described elsewhere.…”

Section: Methodsmentioning

confidence: 99%

Identification of lung major GTP-binding protein as Gi2 and its distribution in various rat tissues determined by immunoassay

Asano

Morishita²,

Semba³

et al. 1989

Biochemistry

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Antisera were raised in rabbits against the 40-kDa alpha subunit of bovine lung GTP-binding protein, which were identified as the alpha subunit of Gi2 (Gi2 alpha) by the analysis of the partial amino acid sequence. Antibodies were purified with a Gi2 alpha-coupled Sepharose column and then were passed through a Gi1 alpha-coupled Sepharose column to remove antibodies reactive also with 41-kDa alpha. Purified antibodies reacted with Gi2 alpha, but not with Gi1 alpha, Gi3 alpha, or Go alpha in an immunoblot assay. A sensitive enzyme immunoassay method for the quantification of Gi2 alpha was developed by using these purified antibodies. The assay system consisted of polystyrene balls with immobilized antibody F(ab')2 fragments and the same antibody Fab' fragments labeled with beta-D-galactosidase from Escherichia coli. The minimal detection limit of the assay was 1 fmol, or 40 pg. Samples from various tissues were solubilized with 2% sodium cholate and 1 M NaCl, and the concentrations of Gi2 alpha were determined. Gi2 alpha was detected in all the tissues examined in the rat. The highest concentration was found in platelets and leukocytes when the data were expressed as picomoles per milligram of protein. The spleen, lung, and cerebral cortex contained relatively high levels of Gi2 alpha. In the bovine brain, Gi2 alpha was distributed almost uniformly among the various regions. The concentrations of Gi2 alpha were constant in the rat brain throughout ontogenic development, in contrast with those of Go alpha which were markedly increased with age.

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Section: Methodsmentioning

confidence: 99%

Identification of lung major GTP-binding protein as Gi2 and its distribution in various rat tissues determined by immunoassay

Asano

Morishita²,

Semba³

et al. 1989

Biochemistry

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“…Thus, fluorimetric ELISA would not be advantageous for detection of putative antiscrapie antibody, under conditions where scrapie agent concentrations were limiting. In contrast, fluorimetric ELISA has greater sensitivity than colorimetric ELISA when saturating concentrations of antigen are adsorbed (18,19,22,23).…”

Section: Resultsmentioning

confidence: 99%

Toward Development of Assays for Scrapie-Specific Antibodies

Kasper

Bowman

Stites

et al. 1981

Hamster Immune Responses in Infectious and Oncologic Diseases

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“…6). A recently described enzyme immunoassay is capable of detecting even 1 attomol of ornitliine-5-aminotransferase (108,109). Other assays, for instance for the determination of a^fetoprotein (51,11Ϊ) or digoxin show a detectability which is more than five times lower than that of the corresponding radioimmunoassays (22).…”

Section: Detection Limitsmentioning

confidence: 99%

Enzyme Immunoassays in Clinical Chemistry: Present Status and Trends

Oellerich¹

1980

CCLM

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The most important enzyme immunoassay techniques are described. The enzymes currently used as labels, the methods of coupling them to antigens or haptens and the possible applications of these assays are reviewed. Furthermore, an overview is given of the reliability and practicability of commercially available enzyme immunoassay kits used in clinical chemical laboratories. Special consideration is given to possible interferences, the detection limits and the mechanization of these tests. Various methods for curve-fitting are listed. It is concluded that most of the currently commercially available enzyme immunoassays are suitable for routine application in appropriate centers like clinical chemical laboratories of larger hospitals. The future role of enzyme immunoassays in clinical chemistry is briefly discussed. Enzymimmunotests in der Klinischen Chemie: Gegenwärtiger Stand und Entwicklungstendenzen Zusammenfassung: Die wichtigsten Techniken des Enzymimmunotests werden beschrieben. Es wird eine Übersicht über die gegenwärtig zur Markierung verwendeten Enzyme, die Methoden zu deren Kopplung an Antigene oder Haptene und die Anwendungsmöglichkeiten dieser Tests gegeben. Ferner wird über den gegenwärtigen Stand der Zuverlässigkeit und Praktikabilität von käuflichen Enzymimmunotests berichtet, welche in klinisch-chemischen Laboratorien angewendet werden. Besondere Berücksichtigung finden die möglichen Interferenzen, die Nachweisgrenzen und Verfahren zur Mechanisierung dieser Tests. Verschiedene mathematische Methoden zur Ermittlung der Ergebniswerte sind aufgeführt. Es wird gefolgert, daß die meisten der zur Zeit käuflichen Enzymimmunotests für die routinemäßige Anwendung in geeigneten Zentren, wie z. B. klinisch-chemischen Laboratorien größerer Kliniken, geeignet sind. Die zukünftige Rolle der Enzymimmunotests in der Klinischen Chemie wird kurz diskutiert. Competitive enzyme immunoassay 2.1.1 2.1.2 Immuno-enzymometric assay 2.2 Competitive sandwich assay 2.3 Sandwich antigen assay 2.4.1 2.4.2

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Enzyme Immunoassay in Rapid Progress

Cited by 26 publications

References 3 publications

Identification of lung major GTP-binding protein as Gi2 and its distribution in various rat tissues determined by immunoassay

Identification of lung major GTP-binding protein as Gi2 and its distribution in various rat tissues determined by immunoassay

Toward Development of Assays for Scrapie-Specific Antibodies

Enzyme Immunoassays in Clinical Chemistry: Present Status and Trends

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