用MALDI-TOF质谱和电子光谱研究纳米胰岛素核-铁蛋白的构建技术和特性

小批量制备了电泳纯鲨鱼肝铁蛋白(Liver ferritin of Sphyma zygaena, SZLF). 用透射电子显微镜技术研究SZLF的铁核和蛋白壳亚基解离和重组过程. 用盐酸解离(pH=1.5)和分离膜透析技术制备铁蛋白亚基, 并用酸碱中和方法重组亚基成为脱铁核铁蛋白(apoSZLF), 同时将胰岛素(Insulin, INS)包裹于apoSZLF蛋白壳内, 构建纳米INS核-SZLF. 用电子光谱、MALDI-TOF质谱和SDS-PAGE技术分别揭示了纳米INS核-SZLF分子结构的真实性, 提出铁蛋白亚基包裹INS构建为纳米INS核-铁蛋白的途径.

Construction Techniques and Characteristics in Nanometer Insulin Core-Ferritin Revealed by MALDI-TOF Mass Spectrometry and Electronic Spectrometry

Liver ferritin of Sphyma zygaena(SZLF) with small-scale preparation was done by electrophoresis approach. SZLF consists of iron core, protein shell, and tunnel across its shell. The pathways for dissociation and recombinant both iron core and protein subunits in SZLF were investigated with transmission electron microscopy. Through dissociation(pH=1.5) of hydrochloric acid and dialysis of separation membrane for preparing ferritin subunits, these free subunits were constructed into apoSZLF under the condition of acid-basis neutralization. In this recombinant process, INS was encapsulated with apoSZLF for constructing a nanometer insulin core(NIC) within the protein shell. The experimental evidence for constructing a NIC-SZLF was revealed by electronic spectrometry, MALDI-TOF MS, and SDS-PAGE, respectively, indicating this novel pathway for a NIC-ferritin encapsulated with the protein subunits and insulin. This technology described can be used to supply an important science evidence for constructing a drug carrier with controlling and releasing characteristics in nanometer peptide core-ferritin.