Development of glutamic acid decarboxylase 65 (GAD65) autoantibody assay using biotin-GAD65 fusion protein.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1130255
- author
- Luo, Dong ; Rogers, Chad N ; Steed, Jordan T ; Gilliam, Lisa K ; Hampe, Christiane S and Lernmark, Åke LU
- publishing date
- 2004
- 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
- 2016-04-01 11:36:34
- date last changed
- 2022-04-12 22:35:01
@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}}, doi = {{10.1016/j.jbiotec.2004.03.009}}, volume = {{111}}, year = {{2004}}, }