The Detailed Preparation Process of GAD and GAD-Ab

GADis the target antigen in type I diabetes patients serum. GAD has now become animportant immunological biomarker in early prediction and diagnosis of diabetes.At first, Anti-GAD Ab was found in StiffmanSyndrome patients by Olimena. Then,many experimental and clinical data show that Anti-GAD Ab can be detected inpatients with autoimmune diseases. Here is a brief introduction of GAD and Anti-GADAb.

GAD widely exists in thenervous system, tissues and organs, such as liver, spleen, pancreas andbrain etc. In mammals, there aretwo  isomers of GAD, one is GAD67 (GAD1) and the other one is GAD65(GAD2). They are encoded by alleles on chromosome 2 and10, respectively. The main differences between GAD67 and GAD65 are shownbelow.

1. The different sensitivity of GADs to react with cofactor-52 pyridoxal phosphate PLP. There is no reaction between GAD67 and PLP, but it is opposite  for GAD65. The activity of GAD65 could be immensely increased after activated by PLP. The difference could be used for identifying various stages of neural development.

2. GAD67 and GAD65 have same cDNA in different organizations, but, their amino acids are different. They only share 65% similarity of amino acids sequence. The main difference lies in the  100 amino acids in N terminal.

3. The type and content of GADs are different in different species. For human, GAD67 mainly expressed in the brain, while GAD65 mainly expressed in brain and pancreas. As shown in Fig.1, there are three isoforms of GAD67, one is GAD67, and the other two isoforms are GAD44 and GAD25, respectively. Due to the different expression location, GAD67 and GAD65 are often used for identification of different biological functions.

Anti-GAD Ab is an early immune marker of type Idiabetes. The positive rate in early type I diabetes can reach 38-76%, and mostof the antibodies are against GAD-65. Cloud-Cloud Corp. has developed thequantitative detection ELISA kit (AED245Hu) for Anti-GAD Ab in vitro.
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