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Analytical biotechnology of recombinant peptides and proteins: I. determination of the purity, composition, and structure of human, porcine, and bovine insulins
Authors:N V Sergeev  I V Nazimov  V G Gavrikov  A I Miroshnikov
Institution:(1) Shemyakin—Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117871 Moscow, GSP-7, Russia;(2) State Scientific Center of Applied Microbiology, 142279 Obolensk, Moscow oblast, Russia
Abstract:A method for analysis of the type, purity, and possible structural modifications of insulins of bovine, porcine, and human origin was proposed. It is based on a combination of narrow-bore reversed-phase HPLC and mass spectrometry. The hydrolysis of insulins with highly specific Glu-protease V8 fromStaphylococcus aureus followed by peptide mapping of the hydrolysis products and mass spectrometry of the isolated fragments helps rapidly and reliably localize and identify substitutions of amino acid residues in insulin structure by using insulin samples of less than 1 nmol.
Keywords:analytical biotechnology  insulin  analysis  mass spectrometry  peptide mapping  protease V8 from Staphylococcus aureus  recombinant proteins  analysis  reversed-phase HPLC
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