TY - JOUR
AU - Yasukawa, Kiyoshi
AB - Abstract Detection of serum autoantibodies to glutamic acid decarboxylase (GAD) is a new method to differentiate insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NTDDM). We established a transformed mouse myeloma cell line, SPG14, which expresses recombinant human GAD65, a major isomer of 65 kDa, inside the cells. GAD65 was partially purified by affinity chromatography using the mouse anti-GAD antibody (Ab). We also established a sandwich ELISA for anti-GAD Ab of the IgG class using GAD65 for coating and the anti-human IgG for detection and examined 54 sera of the IDDM patients and 45 sera of normal individuals. When the mean+2 SD of the color development of the sera of normal individuals was used as a cut-off level, 59.2% of patients with DDDM were positive. This indicates that the ELISA was effective to differentiate D3DM and NDDDM. The result also indicates that the autoantibody to GAD does not dissociate from the recombinant GAD rapidly, even when unbound anti-GAD Ab was fully removed. By using a perfusion culture system, we obtained as many as 4.2 ×1010 SPG14 cells, from which 5 mg of GAD65 could be obtained; this is sufficient for 5,000 assays. This system could be clinically useful for large-scale diagnosis of IDDM. autoantibody, ELISA, GAD, IDDM, myeloma cell © 1997 BY THE JOURNAL OF BIOCHEMISTRY
TI - Expression of Recombinant Human Glutamic Acid Decarboxylase (GAD) in Myeloma Cells and Enzyme-Linked Immunosorbent Assay (ELISA) for Autoantibodies to GAD
JF - The Journal of Biochemistry
DO - 10.1093/oxfordjournals.jbchem.a021563
DA - 1997-01-01
UR - https://www.deepdyve.com/lp/oxford-university-press/expression-of-recombinant-human-glutamic-acid-decarboxylase-gad-in-9ejhrHtmnt
SP - 20
EP - 24
VL - 121
IS - 1
DP - DeepDyve
ER -