Discussion of the methods

Abstract

The insulin RIA was in very great demand right from its establishment and has remained so. The production of antibodies appeared to be very simple, no coupling of the antigen was required, and the first choice of experimental animal, the guinea pig, was fortunate. Minor variations in the immunization procedure regarding dose, frequency, adjuvant, species, purity and formulation of the antigen have been described, but are of slight significance, since it is possible to achieve acceptable antibodies of high capacity and high affinity by immunizing guinea pigs monthly with insulin^50,143. Besides the capacity of the antibodies, very little characterization is normally considered important, except calculating the detection limit (the lowest amount of insulin which can be distinguished from the zero point, p<0.01) and the reproducibility (the day-to-day variation).