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卵清蛋白与多层脂质体及特异性抗卵清蛋白抗体相互作用的自旋标记研究

Spin-labelling study of interactions of ovalbumin with multilamellar liposomes and specific anti-ovalbumin antibodies.

作者信息

Brgles Marija, Mirosavljević Krunoslav, Noethig-Laslo Vesna, Frkanec Ruza, Tomasić Jelka

机构信息

Institute of Immunology Inc., PO Box 266, 10001 Zagreb, Croatia.

出版信息

Int J Biol Macromol. 2007 Mar 10;40(4):312-8. doi: 10.1016/j.ijbiomac.2006.08.012. Epub 2006 Aug 30.

Abstract

Ovalbumin (OVA) has been used continuously as the model antigen in numerous studies of immune reactions and antigen processing, very often encapsulated into liposomes. The purpose of this work was to study the possible interactions of spin-labelled OVA and lipids in liposomal membranes using electron spin resonance (ESR) spectroscopy. OVA was covalently spin-labelled with 4-maleimido-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO-maleimide), characterized and encapsulated into multilamellar, negatively charged liposomes. ESR spectra of this liposomal preparation gave evidence for the interaction of OVA with the lipid bilayers. Such an interaction was also evidenced by the ESR spectra of liposomal preparation containing OVA, where liposomes were spin-labelled with n-doxyl stearic acids. The spin-labelled OVA retains its property to bind specific anti-OVA antibodies, as shown by ESR spectroscopy, but also in ELISA for specific anti-OVA IgG.

摘要

卵清蛋白(OVA)在众多免疫反应和抗原加工研究中一直被用作模型抗原,且常常被包裹在脂质体中。这项工作的目的是利用电子自旋共振(ESR)光谱研究自旋标记的OVA与脂质体膜中脂质之间可能的相互作用。OVA与4-马来酰亚胺-2,2,6,6-四甲基哌啶-1-氧基(TEMPO-马来酰亚胺)共价自旋标记,进行表征后包裹到多层带负电荷的脂质体中。这种脂质体制剂的ESR光谱证明了OVA与脂质双层之间的相互作用。含有OVA的脂质体制剂的ESR光谱也证明了这种相互作用,其中脂质体用n-氧基硬脂酸进行自旋标记。如ESR光谱所示,但在针对特异性抗OVA IgG的ELISA中也表明,自旋标记的OVA保留了其结合特异性抗OVA抗体的特性。

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