Preparation and physical-chemical characterization of poly-N-vinylpyrrolidone-insulin

In an attempt to modify the structure of insulin, the hormone was coupled by covalent linkage to poly-N-vinylpyrrolidone (PVP). PVP-insulin is soluble in aqueous solution and was purified by column chromatography. Special care was taken to check for a possible leakage of insulin from the synthetic polymer. The amount of bound insulin was determined by quantitative amino acid analysis after acid hydrolysis. After protection of the N-terminal glycine residue of the A-chain by citraconylation, a biologically active PVP-insulin was obtained. The molecular weight determined by analytical ultracentrifugation and column chromatography was 50 000-60 000. The biological activity of the coupled hormone was between 0.5 and 7% when compared with native insulin in the system in vitro. The immunological activity was about 20-50%.