Analytical biotechnology of recombinant peptides and proteins. I. Determination of the purity, composition, and structure of human, porcine, and bovine insulins

A method for analysis of the type, purity, and possible structural modifica tions of insulins of bovine, porcine, and human origin was proposed. It is based on a combination of narrow-bore reversed-phase HPLC and mass spectrom etry. The hydrolysis of insulins with highly specific Glu-protease V8 from Staphylococcus aureus followed by peptide mapping of the hydrolysis product s and mass spectrometry of the isolated fragments helps rapidly and reliabl y localize and identify substitutions of amino acid residues in insulin str ucture by using insulin samples of less than 1 nmol.