Analysis of Human Insulin Analogues in Vitro, Using Gel Chromatographic Method
The incubation medium and incubated human pancreas were gel chromatographed on the Bio-Gel P-30 column after extraction with acid ethanol. The extracted immunoreactive insulin (IRI) was successfully separated into two peaks at the position of 6000 molecular weight region. These two peaks corresponded to those which were detected in human serum. These findings suggest that the two groups of insulin are directly secreted from human pancreatic tissue. But the incorporated [3H] leucine peak into acid-ethanol extractable protein did not elute out at the same position as each insulin peak. Therefore, the measurement of [3H] leucine incorporation into acid-ethanol extractable protein is not a good indicator to evaluate insulin biosynthesis.
