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  • 1
    ISSN: 1432-0428
    Keywords: Insulin autoantibodies ; IgG isotype ; light chain ; species specificity ; blood donors ; affinity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Serum samples of 2200 blood donors were screened for anti-insulin IgG by enzyme-linked immunosorbent assay. Specificity of detected antibodies was verified by competition with an excess of insulin and observation that saturated anti-insulin IgG were no longer measurable. The upper limit of measured signal in the population was defined as the mean plus three SD. In the direct assay, 32 sera were positive. Among these, 22 (1%) contained saturable insulin antibodies (true positive) and 10 were non-saturable and considered as non-insulin-specific. The positive blood donors were requested to answer a questionnaire and according to their answers, none had ever received insulin, was a first degree relative of a Type 1 (insulin-dependent) diabetic patient or had experienced a hypoglycaemic episode. None of the 22 true positive sera detected by enzyme-immunosorbent assay bound 125-I-insulin to any significant extent. The nine sera yielding the highest signal were further characterized with regard to heavy and light chains as well as species specificity of ligand. Anti-insulin IgG from healthy blood donors contained only one heavy (γ1 2/9; γ3 7/9) and one light (κ 8/9; λ 1/9) chain. Three sera were human insulin specific; two were non-species-specific; the other four bound insulin in the order human = porcine 〉 bovine. These results indicate that low affinity clonally restricted antibodies against insulin are present in unselected blood donors with a prevalence of 1%.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0428
    Keywords: Anti-insulin ; IgG sub-classes ; monoclonal antibodies ; enzyme linked immunosorbant assay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary An immunochemical micromethod was designed to estimate total IgG and IgG sub-classes of anti-insulin antibodies in immunized diabetic patients. Insulin, immobilized on a solid phase, was allowed to react with serum samples containing anti-insulin antibodies. Bound anti-insulin IgG interacted with mouse monoclonal antibodies specific for total IgG or for each IgG isotype. The fixation of mouse monoclonal antibody was subsequently detected using a horseradish peroxidase-conjugated rabbit anti-mouse IgG in the presence of a chromogenic substrate. The test was standardized by an immunocapture assay utilizing coated rabbit anti-human IgG and known concentrations of purified human myelomatous proteins of each sub-class. Results of anti-insulin IgG and anti-insulin IgG sub-classes assay could therefore be expressed in ng equivalent myelomatous proteins per ml of serum. Analysis of serum samples from 24 insulin-immunized diabetic patients revealed a quasi absence of IgG2 anti-insulin antibodies and an increase of the relative abundance of the other three anti-insulin IgG isotypes. In our series, anti-insulin IgG1 was predominant, followed by IgG3 (in 17/24 patients) or IgG4 (in 7/24). Insulin immunization was deduced to be of polyclonal nature, the isotype pattern of which is not representative of the relative proportion of IgG sub-classes in whole normal serum.
    Type of Medium: Electronic Resource
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