Detection of GAD65 autoantibodies of type-1 diabetes using anti-GAD65-abs reagent produced from bovine brain tissue
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-anti GAD65-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.
Author: Djoko W. Soeatmadji,
Aulanni'am Aulanni'am, Fatchiyah Fatah, Sutiman B. Sumitro
Journal Code: jpkedokterangg050136
