重组胰岛新生相关蛋白（rINGAP）的生产与特性分析
Production and characterization of the recombinant Islet Neogenesis Associated Protein (rINGAP).
作者信息
Assouline-Thomas Béatrice, Pilotte Amélie, Petropavlovskaia Maria, Makhlin Julia, Ding Jieping, McLeod David, Hanley Stephen, Massie Bernard, Rosenberg Lawrence
机构信息
Department of Surgery, McGill University, Montréal, Que., Canada H3G1A4.
出版信息
Protein Expr Purif. 2010 Jan;69(1):1-8. doi: 10.1016/j.pep.2009.07.009. Epub 2009 Jul 25.
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 100x the bioactivity of INGAP peptide on a molar basis in an in vitro assay of human islet regeneration.
胰岛新生相关蛋白（INGAP）与胰岛新生有关。INGAP肽是一种由104 - 118位氨基酸组成的十五肽，可逆转啮齿动物的糖尿病状态，并改善糖尿病患者的葡萄糖稳态。INGAP的作用机制尚不清楚，但此类研究将受益于全长重组蛋白（rINGAP）的可得性。在此，我们报告了慢病毒转导后从293 - SF细胞中产生rINGAP，并通过基质辅助激光解吸电离飞行时间质谱（MALDI - TOF）、四极杆飞行时间质谱（Q - TOF）和高效液相色谱（HPLC）对其进行表征。重要的是，我们发现在人胰岛再生的体外试验中，rINGAP在摩尔基础上表现出比INGAP肽高100倍的生物活性。
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