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Titolo:
Fusion proteins for combined analysis of autoantibodies to the 65-kDa isoform of glutamic acid decarboxylase and islet antigen-2 in insulin-dependentdiabetes mellitus
Autore:
Rickert, M; Seissler, J; Dangel, W; Lorenz, H; Richter, W;
Indirizzi:
Univ Heidelberg, Dept Orthoped Surg, D-96118 Heidelberg, Germany Univ Heidelberg Heidelberg Germany D-96118 , D-96118 Heidelberg, Germany Diabet Res Inst, D-40225 Dusseldorf, Germany Diabet Res Inst Dusseldorf Germany D-40225 , D-40225 Dusseldorf, Germany Labor Dr Koch & Dr Merk, D-88476 Ochsenhausen, Germany Labor Dr Koch & Dr Merk Ochsenhausen Germany D-88476 hsenhausen, Germany
Titolo Testata:
CLINICAL CHEMISTRY
fascicolo: 5, volume: 47, anno: 2001,
pagine: 926 - 934
SICI:
0009-9147(200105)47:5<926:FPFCAO>2.0.ZU;2-#
Fonte:
ISI
Lingua:
ENG
Soggetto:
SERUM-EXCHANGE-WORKSHOP; TYROSINE-PHOSPHATASE; CELL ANTIBODIES; MONOCLONAL-ANTIBODIES; IA-2; PREDICTION; EPITOPES; IDENTIFICATION; AUTOANTIGEN; SPECIFICITY;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
34
Recensione:
Indirizzi per estratti:
Indirizzo: Richter, W Stiftung Orthopad Univ Klin Heidelberg, Schlierbacher Landstr 200A, D-69778 Heidelberg, Germany Stiftung Orthopad Univ Klin Heidelberg Schlierbacher Landstr 200A Heidelberg Germany D-69778
Citazione:
M. Rickert et al., "Fusion proteins for combined analysis of autoantibodies to the 65-kDa isoform of glutamic acid decarboxylase and islet antigen-2 in insulin-dependentdiabetes mellitus", CLIN CHEM, 47(5), 2001, pp. 926-934

Abstract

Background: Prediction, risk assessment, and diagnosis of autoimmune diseases often rely on detection of autoantibodies directed to multiple target antigens, such as the 65-kDa isoform of glutamic acid decarboxylase (GAD65-abs) and the tyrosine phosphatase-like protein islet antigen-2 (IA2-abs), the two major subspecificities of islet cell antibodies (ICAs) associated with insulin-dependent diabetes mellitus, We hypothesized that a combination of autoantigens in a fusion protein unifying the important immunodominant epitopes could provide an efficient target for cost-effective, one-step screening of sera. Methods: Chimeric proteins composed of GAD65 and IA2 residues were constructed, analyzed for their immune reactivity with monoclonal antibodies and sera, and used in a diagnostic assay with S-35-labeled protein as antigen,Results: Length and order of GAD65 and IA2 sequences were critical for conservation of the conformational epitopes in the fusion protein, among four chimera tested, only IA2((606-979))/GAD65((1-585)) retained wild-type-like folding of GAD65 and IA2 domains and yielded a stable protein after baculovirus expression. Reactivity of GAD65 antibody- and IA2 antibody-positive sera from patients newly diagnosed with insulin-dependent diabetes mellitus, from ICA-positive prediabetics, and from ICA-positive first-degree relatives demonstrated conservation of the relevant autoreactive epitopes, The assay based on the in vitro translated fusion antigen had a sensitivity and specificity identical to those for detection of GAD65- and IA2-abs based on the two separate GAD65 and IA2 proteins,Conclusions: Autoantigens such as GAD65 and IA2 can be combined successfully in a fusion protein of similar immune reactivity. This allows simultaneous detection of GAD65- and IA2-abs in a one-step screening assay and cost-effective identification of positive individuals at risk of diabetes or at onset of disease. (C) 2001 American Association for Clinical Chemistry.

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Documento generato il 15/01/21 alle ore 23:03:38