Attempts to prepare 'insulin-free plasma' from human subjects: effects of serial dilution and insulin recovery studies on immunoreactive insulin activity of partially insulin-free plasma.

The reason for attempts to prepare insulin-free plasma from human plasma is discussed. Binding agent, employed to prepare insulin-free plasma, had a capacity to bind 40% of insulin in the system. Insulin recovery from the filtrate of human standard insulin-binding agent reaction was 60%. Human plasma incubated with binding agent gave a filtrate--partially insulin-free plasma (PIFP)--which contained significantly higher amounts of insulin compared to calculated values. Insulin in PIFP was not augmented by serial dilution. Human standard insulin added to PIFP was recovered in full and the mixture was not dilution augmentable. Incubation of human plasma--human standard insulin (106 microunits/ml) mixture with binding agent gave a PIFP which had 60% of total insulin in the mixture. Dialysis did not alter the immunoreactive insulin activity of plasma. Dialysed samples showed augmentation of insulin activity on serial dilution. These observations strongly suggest the presence of a high molecular-weight, insulin-like inhibitory component in human plasma.