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Development of glutamic acid decarboxylase 65 (GAD65) autoantibody assay using biotin-GAD65 fusion protein.

Luo, Dong; Rogers, Chad N; Steed, Jordan T; Gilliam, Lisa K; Hampe, Christiane S and Lernmark, Åke LU (2004) In Journal of Biotechnology 111(1). p.97-104
Abstract
We evaluated a biotin-glutamic acid decarboxylase 65 (GAD65)-based enzyme-linked immunosorbent assay (B-ELISA) to detect GAD65 autoantibodies (GAD65Ab) in 78 sera from individuals with newly diagnosed type 1 diabetes. The GAD65Ab index of patients with type 1 diabetes (mean value of GAD65Ab index of 1.891) was significantly higher than those in 50 sera from healthy control group (mean value of 0.068). The intra- and inter-assay coefficients of variation (CV) were calculated to be 1.042 and 10.703%, respectively. The specificity of the B-GAD65 ELISA was comparable to the standard radioimmunoassay (RIA) which is routinely used in the laboratory. We describe the optimal conditions of the binding kinetics from each assay-step for the detection... (More)
We evaluated a biotin-glutamic acid decarboxylase 65 (GAD65)-based enzyme-linked immunosorbent assay (B-ELISA) to detect GAD65 autoantibodies (GAD65Ab) in 78 sera from individuals with newly diagnosed type 1 diabetes. The GAD65Ab index of patients with type 1 diabetes (mean value of GAD65Ab index of 1.891) was significantly higher than those in 50 sera from healthy control group (mean value of 0.068). The intra- and inter-assay coefficients of variation (CV) were calculated to be 1.042 and 10.703%, respectively. The specificity of the B-GAD65 ELISA was comparable to the standard radioimmunoassay (RIA) which is routinely used in the laboratory. We describe the optimal conditions of the binding kinetics from each assay-step for the detection of GAD65Ab using the WHO standard serum 97/550 as a model autoantibody serum. We concluded that incubation times of 15, 90, and 90 min for step 1 (pre-incubation of Biotin14-GAD65 with serum), step 2 (binding the Ab/Ag complex to NeutrAvidin plate) (Less)
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author
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biotechnology
volume
111
issue
1
pages
97 - 104
publisher
Elsevier
external identifiers
  • scopus:2942519371
ISSN
1873-4863
DOI
10.1016/j.jbiotec.2004.03.009
language
English
LU publication?
no
id
4d2008ef-9c45-4ad7-a477-51518be339f5 (old id 1130255)
date added to LUP
2008-06-16 16:24:16
date last changed
2017-01-01 04:22:02
@article{4d2008ef-9c45-4ad7-a477-51518be339f5,
  abstract     = {We evaluated a biotin-glutamic acid decarboxylase 65 (GAD65)-based enzyme-linked immunosorbent assay (B-ELISA) to detect GAD65 autoantibodies (GAD65Ab) in 78 sera from individuals with newly diagnosed type 1 diabetes. The GAD65Ab index of patients with type 1 diabetes (mean value of GAD65Ab index of 1.891) was significantly higher than those in 50 sera from healthy control group (mean value of 0.068). The intra- and inter-assay coefficients of variation (CV) were calculated to be 1.042 and 10.703%, respectively. The specificity of the B-GAD65 ELISA was comparable to the standard radioimmunoassay (RIA) which is routinely used in the laboratory. We describe the optimal conditions of the binding kinetics from each assay-step for the detection of GAD65Ab using the WHO standard serum 97/550 as a model autoantibody serum. We concluded that incubation times of 15, 90, and 90 min for step 1 (pre-incubation of Biotin14-GAD65 with serum), step 2 (binding the Ab/Ag complex to NeutrAvidin plate)},
  author       = {Luo, Dong and Rogers, Chad N and Steed, Jordan T and Gilliam, Lisa K and Hampe, Christiane S and Lernmark, Åke},
  issn         = {1873-4863},
  language     = {eng},
  number       = {1},
  pages        = {97--104},
  publisher    = {Elsevier},
  series       = {Journal of Biotechnology},
  title        = {Development of glutamic acid decarboxylase 65 (GAD65) autoantibody assay using biotin-GAD65 fusion protein.},
  url          = {http://dx.doi.org/10.1016/j.jbiotec.2004.03.009},
  volume       = {111},
  year         = {2004},
}