Immunogenicity of Synthetic Angiotensin II Octapeptide and Analysis of Antibodies by Passive Haemagglutination

Dietrich, Felix M.; Frischknecht, Hannes

doi:10.1159/000230153

Cited by 7 publications

(2 citation statements)

References 34 publications

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“…Immunochemical analysis of anti¬ bodies to low molecular weight substances exhibiting biological activity (Butler 8c Beiser 1973) may allow comparisons to be drawn between chemical struc¬ tures determining immunological specificity and biological activity (Dietrich Sc Frischknecht 1968). Finally immunological studies of biologically active substances may be useful for their determination of these substances in body fluids or tissue extracts.…”

Section: Discussionmentioning

confidence: 99%

Synthetic Human Calcitonin: Analysis of Antibodies Obtained From Various Animal Species and Determination of Immunoreactive Hormone in Human Sera

Dietrich

Hunziker

Fischer

1975

Acta Endocrinologica

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Antibodies to synthetic human calcitonin (hCT) were developed in rabbits, goats and mice. The free peptide (32 amino-acid residues, Mwt. 3418) was administered together with adjuvant, and the effect of various immunization procedures, as well as of different dose-levels, was evaluated comparatively. Synthetic hCT was found to be a good immunogen for the three animal species examined. The relative importance of various structural parts of the hCT molecule with regard to immunological specificity was determined by reference to the inhibition of the specific binding of 125I-hCT to antibodies by peptide fragments of hCT.All the antisera studied were directed to structural and/or conformational properties of the 11\p=n-\28 or 11\p=n-\32amino acid sequence of hCT. Six different antisera from rabbits and goats were selected for radioimmunological assay of hCT on the basis of their inhibitory dose50-values and immunological specificity. To improve the sensitivity of the radioimmunoassay (RIA), we studied the preparation of radioiodinated hCT and assessed various parameters determining the sensitivity of the assay. Despite all the efforts, CT in human plasma from healthy subjects could not be determined with certainty. The difficulties encountered in the

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Section: Discussionmentioning

confidence: 99%

Synthetic Human Calcitonin: Analysis of Antibodies Obtained From Various Animal Species and Determination of Immunoreactive Hormone in Human Sera

Dietrich

Hunziker

Fischer

1975

Acta Endocrinologica

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“…In the few cases in which the sandwich assay format is used for hapten detection, the second binding is achieved by an antimetatype antibody that recognizes the haptenantibody complex (8 -9 ), by an antiidiotype antibody such as ␣-AId (10,11 ), or by the labeled VH (heavy chain variable domain) chain in the open sandwich assay format (12,13 ). Until now, the smallest natural molecule for which a true sandwich assay has been reported was angiotensin II, a linear nonhapten (14 ) octapeptide having a molecular mass of 1048 Da. The 2 antibodies used for this sandwich assay were raised against the molecular fragments of the C and N termini, respectively (15 ).…”

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confidence: 99%

Sandwich Assay for Tacrolimus Using 2 Antitacrolimus Antibodies

et al. 2014

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BACKGROUND Although detection of natural haptens by antihapten antibodies in sandwich assay format has the theoretical advantages of high analytical specificity and sensitivity, this type of assay has not been reported because of the seemingly insurmountable task of avoiding steric hindrance between the 2 bindings. This is especially true for ring-structured hydrophobic haptens. The macrolide drug tacrolimus (FK506, Prograf®, 804 Da) is such a hapten. Here we show the detection of tacrolimus using 2 antitacrolimus monoclonal antibodies in a sandwich assay. METHODS Both antibodies were developed by use of an intact tacrolimus molecule covalently linked to a carrier protein but via 2 different positions separated by 10 carbon atoms. Epitope analysis based on drug analog binding was used to show no overlap between the binding sites of the 2 antibodies, indicating the 10-carbon separation resulted in 2 distinct epitopes. The distinct epitopes suggested that the drug might be approachable by the antibodies from 2 separate directions, which predicted simultaneous binding as in sandwich formation. RESULTS This prediction was confirmed in sandwich ELISA and affinity column–mediated immunoassay formats. The assay demonstrated good imprecision and significantly lower metabolite cross-reactivity than competitive assay counterparts. Comparison with liquid chromatography–tandem mass spectrometry (LC-MS/MS) using 55 whole-blood samples from transplant patients with tacrolimus concentrations ranging from 0.9 to 29.5 ng/mL showed a linear regression: sandwich = 0.99 × LC-MS/MS + 0.10 ng/mL, r = 0.991, Sy|x = 1.08 ng/mL. CONCLUSIONS This work demonstrates that a highly specific sandwich assay using 2 antihapten antibodies is feasible for the measurement of a hapten drug.

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Immunogenicity and Antigenicity of Angiotensin I and II

Vallotton

1974

Angiotensin

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Immunogenicity of Synthetic Angiotensin II Octapeptide and Analysis of Antibodies by Passive Haemagglutination

Cited by 7 publications

References 34 publications

Synthetic Human Calcitonin: Analysis of Antibodies Obtained From Various Animal Species and Determination of Immunoreactive Hormone in Human Sera

Synthetic Human Calcitonin: Analysis of Antibodies Obtained From Various Animal Species and Determination of Immunoreactive Hormone in Human Sera

Sandwich Assay for Tacrolimus Using 2 Antitacrolimus Antibodies

Immunogenicity and Antigenicity of Angiotensin I and II

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