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- Title
Preparation and Testing of Diabetes Autoantibody Controls.
- Authors
Brooking, H.; Powell, M.J.; Amoroso, M.; Betterle, C.; Pedini, B.; Coco, G.; Furmaniak, J.; Smith, B. Rees
- Abstract
Stocks of the WHO islet cell antibody, GAD65 antibody, and IA-2 antibody standard (NIBSC 97/550) are now very limited. We have therefore made and tested a series of control preparations in which human monoclonal autoantibodies to IA-2 and to GAD65 were diluted in antibody-negative human serum to different concentrations. Three different diabetes autoantibody controls (DAC 1–3) were made as was a negative control preparation. Aliquots containing 1 mL of autoantibodies in serum were freeze-dried. After reconstitution (with 1 mL of water) the controls were tested by 125I-IA-2 immunoprecipitation assay (IPA), 125I-GAD65 IPA, GAD65 Ab ELISA and IA-2 Ab ELISA (kits from RSR Ltd.) and for ICA by immunofluorescence test (IFT). DAC1 is particularly suitable as a control for the 125I IA-2 IPA; DAC2 is suitable for the 125I-GAD65 IPA, GAD65 Ab ELISA, and IA-2 Ab ELISA; and DAC3 is suitable for the ICA IFT. Freeze-dried preparations showed good stability at 37°C. Reconstituted liquid preparations were stable when stored at 4°C and at 37°C. Availability of an essentially unlimited supply of these reagents should be useful in establishing reproducible and comparable measurements of diabetes autoantibodies in different laboratories using different assays.
- Subjects
DIABETES; AUTOANTIBODIES; MONOCLONAL antibodies; ISLANDS of Langerhans; IMMUNOFLUORESCENCE; MEDICAL research
- Publication
Annals of the New York Academy of Sciences, 2008, Vol 1150, p316
- ISSN
0077-8923
- Publication type
Article
- DOI
10.1196/annals.1447.014