A Quantitative Theory of the Precipitin Reaction

Stokinger, Herbert E.; Heidelberger, Michael

doi:10.1084/jem.66.2.251

Cited by 60 publications

(14 citation statements)

References 13 publications

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“…Similarly, the reactivity of Abs produced by the transformed cells may also be affected, with the cells derived from the sickest patients producing a higher frequency of Abs that react nonspecifically or "polyspecifically" (33) with HIV compared to the Abs from cells derived from patients in whom the disease process is more limited ( Table 2). Alternatively, the degree of true cross-reactivity between anti-HIV Abs and various mammalian proteins may increase with length of exposure to HIV, recapitulating the studies of Stokinger and Heidelberger (34) showing proportional increases in cross-reacting Abs with prolonged immunization. This association between disease progression and the decrease in efficiency with which EBV can infect B cells as well as the increase in production of nonspecific, polyspecific, or cross-reacting Abs may reflect the polyclonal activation of B cells and is thought to contribute to the immunosuppression in HIV-infected individuals (35) and to many of the clinical manifestations of the disease (36).…”

Section: Resultsmentioning

confidence: 80%

Generation of human monoclonal antibodies to human immunodeficiency virus.

Górny

Gianakakos

Sharpe

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

170

127

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Section: Resultsmentioning

confidence: 80%

Generation of human monoclonal antibodies to human immunodeficiency virus.

Górny

Gianakakos

Sharpe

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

170

127

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“…This was done by mixing the antiserum with human serum in the proportion of 1 to 0.25, incubating the mixture overnight, and centrifuging the small precipitate formed. A similar absorption technique was used by Stokinger and Heidelberger (5).…”

Section: Methodsmentioning

confidence: 99%

Iodine Components of the Blood. Circulating Thyroglobulin in Normal Persons and in Persons With Thyroid Disease

Jacob¹

1940

J. Clin. Invest.

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It is now well established that the amount of iodine circulating in the blood is roughly an index of thyroid activity. Iodine values below 5 gamma per cent are usually found in myxedema, and values over 10 gamma per cent are suggestive of hyperthyroidism. A portion of the blood iodine is inorganic and presumably inert in a hormonal sense; the remainder is organic and probably represents the circulating hormone or its components. Very little is known of the nature of this organic iodine. In the course of experimfients on the production of antibodies to human thyroglobulin, it seemed that antiserum potent with respect to thyroglobulin antibodies might be used to detect thyroglobulin in human serum by appropriate immunologic reactions. METHODThe technique used to obtain antithyroglobulin serum is similar to that of Hektoen and Schulhof (1), Rosen and Marine (2), and Schulhof (3). Thyroglobulin was prepared from human thyroid glands which had been removed at operation and made relatively free from serum.1 It was kept in suspension at its isoelectric point. The protein content was usually 1.5 to 2.0 per cent, and the iodine about 5 to 6 mgm. per cent. This material was injected into rabbits intraperitoneally, intravenously, or into subcutaneous nodules according to the method described by Dienes (4). The injections were given for 2 to 3 days in succession, with rest periods of 4 to 6 days. The intraperitoneal injection contained about 75 to 150 mgm. of thyroglobulin, the intravenous injection 15 to 20 mgm. and the intranodular injection 1 to 3 mgm. for each nodule. After a period of 4 to 8 weeks, the serum of such animals usually contained sufficient antibodies to be of value in testing for very small amounts of thyroglobulin. In some instances, animals were injected for several months in succession in order to increase their antibody titer. The rabbit antiserum contained not only antibodies for human thyroglobulin but also small amounts of antibodies for human serum protein. before using such serum in immunologic tests it was necessary to absorb the antibodies against human serum. This was done by mixing the antiserum with human serum in the proportion of 1 to 0.25, incubating the mixture overnight, and centrifuging the small precipitate formed. A similar absorption technique was used by Stokinger and Heidelberger (5).The presence of antibodies was determined by the ring precipitin test, using the undiluted rabbit serum against dilutions of thyroglobulin or human serum. Thyroglobulin was first dissolved in dilute alkali at a pH of 8.0 to 9.0 until it was almost clear and the insoluble portion removed by centrifugation. Normal saline used in making dilutions was also adjusted to the same pH. Otherwise, at the usual pH of normal saline there would be precipitation of thyroglobulin.2 The antiserum was introduced at the bottom of small tubes and the dilutions of thyroglobulin or human serum were layered on top. The results were read after 1 to 2 hours at room temperature.In attempting to evaluate the potency of the anti...

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“…In a retrospective assessment of the utility of multiple potential markers of the recurrence of TC, post-treatment Tg concentration was found to be the strongest independent predictor of the recurrence (3). The presence of endogenous anti-Tg autoantibodies (Tg-AAb) can mask the epitopes used by reagent antibodies in immunoassay for measurement of Tg, which can lead to falsely negative results (4–7). Tg autoantibodies were first described by Stokinger and Heidelberger (4); active research of Tg-AAb began over 50 years ago (8), but to date there are no commercially distributed immunoassays available that can overcome the interference of Tg-AAb in testing for Tg.…”

Section: Introductionmentioning

confidence: 99%

Measurement of Thyroglobulin by Liquid Chromatography–Tandem Mass Spectrometry in Serum and Plasma in the Presence of Antithyroglobulin Autoantibodies

Kushnir

Rockwood

Roberts³

et al. 2013

Clinical Chemistry

170

119

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Background Measurement of serum thyroglobulin (Tg) is used to monitor patients after treatment for differentiated thyroid carcinoma (TC). Difficulty in using Tg as a biomarker of the recurrence of TC in many patients stems from the presence of endogenous anti-Tg autoantibodies (Tg-AAb), which can interfere with immunoassays (IA) and cause false-negative results. Methods Tg was enriched from serum samples using rabbit polyclonal anti-Tg antiserum and protein precipitation. Unrelated proteins were partially depleted in the process. Enriched proteins were then denatured, reduced, and digested with trypsin after the addition of a winged internal standard peptide. A Tg-specific tryptic peptide was purified by immunoaffinity extraction and analyzed by 2D-LC-MS/MS. Instrument cycle time was 6.5 min per sample. Results The lower limit of quantification was 0.5 ng/mL (0.76 fmol/mL of dimer). Total imprecision of triplicate measurements in serum samples over five days was less than 10%. Comparison with a commercial IA using serum samples free of Tg-AAb (n=73) showed Deming regression, IA= 1.00*LC-MS/MS-2.35, r=0.982, Sy,x=9.52. In a set of Tg-AAb positive samples tested negative for Tg using IA (n=71), concentrations determined by LC-MS/MS method were at or above 0.5 in 23% of samples (median 1.2, range 0.7–11 ng/mL). Conclusions The method has acceptable performance characteristics for use in clinical diagnostic applications. The most substantial disagreement between the methods was observed in Tg-AAb positive samples with concentration below 2 ng/mL (determined with LC-MS/MS). The affinity assisted enrichment strategy used for Tg in this method is applicable to other biomarkers that have endogenous autoantibodies.

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A Quantitative Theory of the Precipitin Reaction

Cited by 60 publications

References 13 publications

Generation of human monoclonal antibodies to human immunodeficiency virus.

Generation of human monoclonal antibodies to human immunodeficiency virus.

Iodine Components of the Blood. Circulating Thyroglobulin in Normal Persons and in Persons With Thyroid Disease

Measurement of Thyroglobulin by Liquid Chromatography–Tandem Mass Spectrometry in Serum and Plasma in the Presence of Antithyroglobulin Autoantibodies

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