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Determination of glutamic acid decarboxylase antibodies (GADA) IgG subclasses - comparison of three immunoprecipitation assays (IPAs)

Hillman, Magnus LU ; Törn, Carina LU and Landin-Olsson, Mona LU (2007) In Clinical and Experimental Immunology 150(1). p.68-74
Abstract
IgG subclasses of glutamic acid decarboxylase (GAD(65)) antibodies (GADA) may reflect the immunological state in the pancreas of GADA-positive patients with autoimmune diabetes. The use of biotin-conjugated antibodies and streptavidin Sepharose are used commonly in immunoprecipitation assays (IPA) based on (125)I- or (35)S-labelled antigens to capture IgG subclasses directed against IA-2 or GAD(65). We have compared three different immunoprecipitation assays for the determination of GADA IgG subclasses. Two of the assays were based on the biotin and streptavidin systems provided in a solid (immobilized) or liquid (mobilized) phase binding environment. The third assay was based on N-hydroxysuccinimide (immobilized) interaction with primary... (More)
IgG subclasses of glutamic acid decarboxylase (GAD(65)) antibodies (GADA) may reflect the immunological state in the pancreas of GADA-positive patients with autoimmune diabetes. The use of biotin-conjugated antibodies and streptavidin Sepharose are used commonly in immunoprecipitation assays (IPA) based on (125)I- or (35)S-labelled antigens to capture IgG subclasses directed against IA-2 or GAD(65). We have compared three different immunoprecipitation assays for the determination of GADA IgG subclasses. Two of the assays were based on the biotin and streptavidin systems provided in a solid (immobilized) or liquid (mobilized) phase binding environment. The third assay was based on N-hydroxysuccinimide (immobilized) interaction with primary amines (i.e. lysine residues) on the antibody. We found the liquid phase binding assay (LPBA) to be the most stable assay, with a comparatively low coefficient of variation and background. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Clinical and Experimental Immunology
volume
150
issue
1
pages
68 - 74
publisher
British Society for Immunology
external identifiers
  • pmid:17666094
  • wos:000249275500008
  • scopus:34548526770
ISSN
0009-9104
DOI
10.1111/j.1365-2249.2007.03473.x
language
English
LU publication?
yes
id
374a8f53-189e-47cc-916f-bec737f3c9fb (old id 1142365)
date added to LUP
2008-08-07 12:21:39
date last changed
2017-01-01 04:32:49
@article{374a8f53-189e-47cc-916f-bec737f3c9fb,
  abstract     = {IgG subclasses of glutamic acid decarboxylase (GAD(65)) antibodies (GADA) may reflect the immunological state in the pancreas of GADA-positive patients with autoimmune diabetes. The use of biotin-conjugated antibodies and streptavidin Sepharose are used commonly in immunoprecipitation assays (IPA) based on (125)I- or (35)S-labelled antigens to capture IgG subclasses directed against IA-2 or GAD(65). We have compared three different immunoprecipitation assays for the determination of GADA IgG subclasses. Two of the assays were based on the biotin and streptavidin systems provided in a solid (immobilized) or liquid (mobilized) phase binding environment. The third assay was based on N-hydroxysuccinimide (immobilized) interaction with primary amines (i.e. lysine residues) on the antibody. We found the liquid phase binding assay (LPBA) to be the most stable assay, with a comparatively low coefficient of variation and background.},
  author       = {Hillman, Magnus and Törn, Carina and Landin-Olsson, Mona},
  issn         = {0009-9104},
  language     = {eng},
  number       = {1},
  pages        = {68--74},
  publisher    = {British Society for Immunology},
  series       = {Clinical and Experimental Immunology},
  title        = {Determination of glutamic acid decarboxylase antibodies (GADA) IgG subclasses - comparison of three immunoprecipitation assays (IPAs)},
  url          = {http://dx.doi.org/10.1111/j.1365-2249.2007.03473.x},
  volume       = {150},
  year         = {2007},
}