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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 LU ; Landin-Olsson, Mona LU ; Ortqvist, E; Palmer, J P and Lernmark, Åke LU , et al. (2005) In Clinical and Experimental Immunology 140(3). p.564-571
Abstract
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... (More)
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. (Less)
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Contribution to journal
publication status
published
subject
keywords
type 1 diabetes, epitopes, insulin autoantibody, radioligand binding, assay, recombinant Fab
in
Clinical and Experimental Immunology
volume
140
issue
3
pages
564 - 571
publisher
British Society for Immunology
external identifiers
  • wos:000229227000021
  • pmid:15932520
  • scopus:20444473243
ISSN
0009-9104
DOI
10.1111/j.1365-2249.2005.02802.x
language
English
LU publication?
yes
id
a28c86a2-6624-4b2d-9d3c-c74f97f13f1b (old id 240287)
date added to LUP
2007-08-10 16:12:20
date last changed
2017-01-01 04:33:08
@article{a28c86a2-6624-4b2d-9d3c-c74f97f13f1b,
  abstract     = {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 &lt; 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.},
  author       = {Padoa, C J and Crowther, N J and Thomas, J W and Hall, T R and Bekris, L M and Törn, Carina and Landin-Olsson, Mona and Ortqvist, E and Palmer, J P and Lernmark, Åke and Hampe, C S},
  issn         = {0009-9104},
  keyword      = {type 1 diabetes,epitopes,insulin autoantibody,radioligand binding,assay,recombinant Fab},
  language     = {eng},
  number       = {3},
  pages        = {564--571},
  publisher    = {British Society for Immunology},
  series       = {Clinical and Experimental Immunology},
  title        = {Epitope analysis of insulin autoantibodies using recombinant Fab},
  url          = {http://dx.doi.org/10.1111/j.1365-2249.2005.02802.x},
  volume       = {140},
  year         = {2005},
}