Simple Method for Development of Sensitive and Specific Antiinsulin Antisera for Laboratory Use
- 1 March 1984
- journal article
- research article
- Published by Taylor & Francis in Journal of Immunoassay
- Vol. 5 (1-2), 131-144
- https://doi.org/10.1080/01971528408063003
Abstract
Commercial sources provide good, though expensive antiinsulin antisera. We describe here a simple, fast and inexpensive method for the production of antiinsulin antisera. Purified pork insulin (Lente) was injected subcutaneously in oil/water/complete Freund adjuvant mixture. Three guinea pigs received 0.25 mg of insulin and three received 0.5 mg of insulin. Subsequent injections of the same dose were done 40 and 60 days later. Five animals developped antisera with titers superior to 1:10,000 40 days after the primary inoculation. Four out of five guinea pigs improved further their antibody titer after the 2nd and 3rd injection (p < 0.0005). Good sensitivity was associated with titers superior to 1:50,000 and appeared only after the 2nd injection to improve further after the 3rd. Thus, four out of six animals developed antiinsulin antisera suitable for the radioimmunoassay (RIA). The antisera bound pro-insulin on an equimolar basis with insulin while glucagon was not bound up to 100 ng/ml. The minimum detectable insulin concentration was about 12 pg (0.3 μU) at the optimum antiserum dilution. Six animals given a small dose of insulin (0.06 mg) developed antisera of a low titer and sensitivity, unsuitable for the RIA.Keywords
This publication has 4 references indexed in Scilit:
- A METHOD FOR PRODUCING SPECIFIC ANTISERA WITH SMALL DOSES OF IMMUNOGENJournal of Clinical Endocrinology & Metabolism, 1971
- Immune response to insulin in guinea pigsMetabolism, 1971
- Monoiodoinsulin: Demonstration of its biological activity and binding to fat cells and liver membranesBiochemical and Biophysical Research Communications, 1971
- Immunological reactivity of insulin to sepharose coupled with insulin-antibody—Its use for the extraction of insulin from serumBiochemical and Biophysical Research Communications, 1970