Detection of GAD65 autoantibodies of type-1 diabetes using anti-GAD65-abs reagent produced from bovine brain tissue

  • Djoko W. Soeatmadji
  • Aulanni'am Aulanni'am
  • Fatchiyah Fatah
  • Sutiman B. Sumitro
Keywords: GAD, type-1 Diabetes
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Abstract

Clinically, type 1 diabetes may presents as type 2 diabetes which sometimes not easily differentiated. Perhaps only autoimmune markers of β-cells destruction could differentiate those two clinical conditions. Due to extremely high cost ( $ 150/test), examination of anti-glutamic acid decarboxylase-65 auto-antibodies (anti-GAD65Abs) may not be routinely performed in most, if not all, clinical laboratories in Indonesia. Hence, the production of anti-GAD65 Abs reagent in Indonesia may reduce the cost and improve the quality of diabetes care in Indonesia. We produce reagent to detect anti-GAD65-Abs using bovine brain tissue as source of GAD enzyme in 3 steps. Step 1, isolation, purification of GAD65 from bovine brain tissue and used it as a primary antigen to stimulate the generation of anti-GAD65 antibodies in Wistar rat. Step 2, the purified GAD65 antibodies were than used as a secondary antibody to induce the production of anti-anti-GAD65-antibodies in Wistar rat and rabbit. Step 3. Labeling anti-antiGAD65-antibodies with alkaline phoshpatase and peroxidase, and detecting anti-GAD65Abs previously detected using commercial kit. The anti-anti-GAD65-antibodies reagent produced in our laboratories successfully identify anti-GAD65-Abs of type 1 diabetic patients previously detected with commercial reagent. (Med J Indones 2005; 14: 197-203)
Published
2005-11-01
How to Cite
1.
Soeatmadji DW, Aulanni’am A, Fatah F, Sumitro SB. Detection of GAD65 autoantibodies of type-1 diabetes using anti-GAD65-abs reagent produced from bovine brain tissue. Med J Indones [Internet]. 2005Nov.1 [cited 2020Dec.3];14(4):197-03. Available from: http://mji.ui.ac.id/journal/index.php/mji/article/view/196
Section
Basic Medical Research

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