Epitope analysis of insulin autoantibodies using recombinant Fab
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Epitope analysis of insulin autoantibodies using recombinant Fab. / Padoa, C J; Crowther, N J; Thomas, J W; Hall, T R; Bekris, L M; Törn, Carina; Landin-Olsson, Mona; Ortqvist, E; Palmer, J P; Lernmark, Åke; Hampe, C S.
I: Clinical and Experimental Immunology, Vol. 140, Nr. 3, 2005, s. 564-571.Forskningsoutput: Tidskriftsbidrag › Artikel i vetenskaplig tidskrift
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T1 - Epitope analysis of insulin autoantibodies using recombinant Fab
AU - Padoa, C J
AU - Crowther, N J
AU - Thomas, J W
AU - Hall, T R
AU - Bekris, L M
AU - Törn, Carina
AU - Landin-Olsson, Mona
AU - Ortqvist, E
AU - Palmer, J P
AU - Lernmark, Åke
AU - Hampe, C S
PY - 2005
Y1 - 2005
N2 - Autoantibodies to insulin are often the first autoantibodies detected in young children with type 1 diabetes and can be present before the onset of clinical diabetes. These autoantibodies and their epitopes are, however, not well characterized. We explored the use of monoclonal antibodies and their recombinant Fab as reagents for epitope analysis. In this study we cloned and characterized the recombinant Fab of the insulin-specific monoclonal antibody CG7C7. We found the epitope of this antibody to be located predominantly at the A-chain loop of the insulin molecule. The recombinant Fab was then used to compete for insulin binding against insulin autoantibodies present in sera from patients with type 1 or type 1.5 diabetes. In competition experiments with sera positive for autoantibodies to insulin the recombinant Fab significantly reduced the binding to [I-125]-insulin by sera of type 1 (n = 35) and type 1.5 diabetes [latent autoimmune diabetes in adults (LADA)] (n = 14) patients (P < 0.0001). We conclude that competition between insulin-specific monoclonal antibodies or their recombinant Fab and insulin autoantibodies should prove useful in the epitope analysis of autoantibodies to insulin.
AB - Autoantibodies to insulin are often the first autoantibodies detected in young children with type 1 diabetes and can be present before the onset of clinical diabetes. These autoantibodies and their epitopes are, however, not well characterized. We explored the use of monoclonal antibodies and their recombinant Fab as reagents for epitope analysis. In this study we cloned and characterized the recombinant Fab of the insulin-specific monoclonal antibody CG7C7. We found the epitope of this antibody to be located predominantly at the A-chain loop of the insulin molecule. The recombinant Fab was then used to compete for insulin binding against insulin autoantibodies present in sera from patients with type 1 or type 1.5 diabetes. In competition experiments with sera positive for autoantibodies to insulin the recombinant Fab significantly reduced the binding to [I-125]-insulin by sera of type 1 (n = 35) and type 1.5 diabetes [latent autoimmune diabetes in adults (LADA)] (n = 14) patients (P < 0.0001). We conclude that competition between insulin-specific monoclonal antibodies or their recombinant Fab and insulin autoantibodies should prove useful in the epitope analysis of autoantibodies to insulin.
KW - type 1 diabetes
KW - epitopes
KW - insulin autoantibody
KW - radioligand binding
KW - assay
KW - recombinant Fab
U2 - 10.1111/j.1365-2249.2005.02802.x
DO - 10.1111/j.1365-2249.2005.02802.x
M3 - Article
VL - 140
SP - 564
EP - 571
JO - Clinical and Experimental Immunology
JF - Clinical and Experimental Immunology
SN - 0009-9104
IS - 3
ER -