Production and characterization of the recombinant Islet Neogenesis Associated Protein (rINGAP) |
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| Authors: | Béatrice Assouline-Thomas Amélie Pilotte Maria Petropavlovskaia Julia Makhlin Jieping Ding David McLeod Stephen Hanley Bernard Massie Lawrence Rosenberg |
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| Institution: | 1. Genetics and Molecular Pathology, SA Pathology, North Adelaide, South Australia, Australia;2. Department of Paediatrics, The University of Adelaide, Adelaide, South Australia, Australia;3. Department of Genetics, The University of Adelaide, South Australia, Australia;4. Mesenchymal Stem Cell Laboratory, School of Medical Sciences, The University of Adelaide, Adelaide, South Australia, Australia;1. College of Pharmacy and Gachon Institute of Pharmaceutical Sciences, Gachon University, Incheon 406-799, Republic of Korea;2. School of Natural Resources and Environmental Sciences, Kangwon National University, Chuncheon 24341, Republic of Korea |
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| Abstract: | Islet Neogenesis Associated Protein (INGAP) is implicated in pancreatic islet neogenesis. INGAP peptide, a pentadecapeptide comprising amino acids 104–118, reverses diabetes in rodents and improves glucose homeostasis in patients with diabetes. The mechanism of INGAP action is unknown, but such studies would benefit from the availability of the full-length recombinant protein (rINGAP).Here we report the production of rINGAP from 293-SF cells following lentiviral transduction, and its characterization by MALDI-TOF and Q-TOF Mass Spectrometry, and HPLC.Importantly, we show that rINGAP exhibits 100× the bioactivity of INGAP peptide on a molar basis in an in vitro assay of human islet regeneration. |
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