The interpretation of paradoxical radioimmunoassay standard curves

Niederer, Werner

doi:10.1016/0022-1759(74)90048-9

Cited by 12 publications

(3 citation statements)

References 9 publications

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“…[14C]proline were purified as described (9,11,14), and 10 ,xg of the collagen dissolved in 0.05M tris-HCl, 0.9M NaCl, and 0.1 percent bovine thyroglobulin was incubated with nitrocellulose filters (Millipore SCWP; 13 mm; pore size, 8 ,um) in a volume of 1 ml at 4°C for 10 hours. Substrate binding efficiency was greater than 90 percent.…”

Section: Collagens Labeled Biosynthetically Withmentioning

confidence: 99%

“…The substrates were thus extracted, purified, and incubated with the nitrocellulose under nondenaturing conditions. The substrates were judged to be native because of their tertiary structure observed by rotary-shadowing electron microscopy and their insensitivity to digestion by ot-thrombin at 30°C (14). The endothelial chemoattractant we used was derived from retina extracts (15).…”

Section: Collagens Labeled Biosynthetically Withmentioning

confidence: 99%

“…Immunoassays have had extensive applications in many fields. Although CIA's with the use of serum (12)(13)(14) have been described, the mechanism of the cooperativity has not been understood. On the basis of the observations of Weintraub et al (13) that the effect is lost when F(ab) fragments are prepared, and on the basis of our findings with monoclonal antibodies (2, 7), we assert that formation of a circular complex may frequently be the cause for cooperativity in antisera.…”

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Cooperative Immunoassays: Ultrasensitive Assays with Mixed Monoclonal Antibodies

Ehrlich¹,

Moyle²

1983

Science

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Mixtures of certain monoclonal antibodies appear to bind human chorionic gonadotropin in a "cooperative" fashion because they form circular complexes with the hormone. Experiments illustrate how this property might be exploited to develop very sensitive immunoassays for human chorionic gonadotropin or any other antigen. Since the assays are not based on competitive inhibition between radiolabeled and unlabeled antigen, they are much more sensitive than a traditional radioimmunoassay in which either one of the same antibodies is used alone.

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Section: Collagens Labeled Biosynthetically Withmentioning

confidence: 99%

Section: Collagens Labeled Biosynthetically Withmentioning

confidence: 99%

mentioning

confidence: 99%

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Cooperative Immunoassays: Ultrasensitive Assays with Mixed Monoclonal Antibodies

Ehrlich¹,

Moyle²

1983

Science

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Interaction of Immobilized Avidin with an Aequorin–Biotin Conjugate: An Aequorin-Linked Assay for Biotin

Feltus

Ramanathan

Daunert

1997

Analytical Biochemistry

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Quantitative immunocytochemistry of pituitary receptors for luteinizing hormone-releasing hormone

1975

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In Araldite sections of male rat pituitaries, stained after embedding by the unlabeled antibody enzyme method with antisera to native luteinizing hormone-releasing hormone (LH-RH) or LH-RH azo-conjugated to bovine serum albumin, localization is confined mainly to the interior of the large, and to a lesser extent to that of the small, secretion granules of the gonadotrophic cells. Plasma membranes are not demonstrated. Except for weak staining in the granules of corticotrophs, no other pituitary cell is stained. Pretreatment of sections with LH-RH (to dilutions of 4 pg/mul) increases staining intensity in the gonadotrophic granules. Other cells are unaffected. The lesser the gonadotroph staining intensity without pretreatment, the greater the increase (more than 23-fold reactivity). Augmented staining is measurable (P less than 0.001) to antiserum dilutions of 1:240000. Pretreatment with des-Glu-1-LH-RH, porcine corticotropin or rat prolactin has no effect. LH-RH-Gly-10(des-amide) inhibits. Rat glycoprotein hormones enhance staining with anti-azo-conjugated LH-RH. With antinative LH-RH these hormones enhance weak staining, but inhibit strong staining. Thick vibrotome sections of male rat or rabbit pituitaries stained before embedding reveal specific localization on plasma membrane and gonadotrophic secretion granules provided the sections have been pretreated with LH-RH (250 pg/mul). The data show that LH-RH after reaction with receptor is not sterically hindered from binding specific antibodies. Receptor may be found in secretion granules, both in the free state or combined with LH-RH. Plasma membrane receptor, on the other hand, was free under the conditions of the experiments. Immunization with LH-RH elicits not only heteroimmune antibodies specific for LH-RH, but also a group of still ill defined autoimmune antibodies, some of which may conceivably be reactive with glycoprotein hormone alpha-chains.

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The interpretation of paradoxical radioimmunoassay standard curves

Cited by 12 publications

References 9 publications

Cooperative Immunoassays: Ultrasensitive Assays with Mixed Monoclonal Antibodies

Cooperative Immunoassays: Ultrasensitive Assays with Mixed Monoclonal Antibodies

Interaction of Immobilized Avidin with an Aequorin–Biotin Conjugate: An Aequorin-Linked Assay for Biotin

Quantitative immunocytochemistry of pituitary receptors for luteinizing hormone-releasing hormone

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