The isolation and purification of biologically active recombinant and native autoantigens for the study of autoimmune disease

被引：33

作者：

Apostolopoulos, J ^{[1
]}

Ooi, JDK ^{[1
]}

Odobasic, D ^{[1
]}

Holdsworth, SR ^{[1
]}

Kitching, AR ^{[1
]}

机构：

[1] Monash Univ, Dept Med, Monash Med Ctr, Ctr Inflammatory Dis, Clayton, Vic 3168, Australia

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2006年 / 308卷 / 1-2期

关键词：

myeloperoxidase; mouse collagen alpha 3 type IV; baculovirus; glomerulonephritis;

D O I：

10.1016/j.jim.2005.10.011

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The expression of recombinant, biologically active mouse myeloperoxidase (MPD) and the recombinant non-collagetious (NC1) domain of mouse collagen alpha 3 Type IV was achieved for the first time in Sf21 cells (Spodoptera frugiperda ovarian insect cells) using a baculovirus expression system. Following purification, the proteins were identified by reducing and nonreducing SDS-PAGE clectrophoresis. Recombinant mouse MPO has a molecular weight of approximately 90 kDa and mouse alpha 3(IV)NC1 approximately 32 kDa. In addition, milligram quantities of native mouse myeloperoxidase were purified from 32Dcl3 cells. Both native and recombinant myeloperoxidase were biologically active. This study also demonstrated that the immunization of myeloperoxidase deficient (Mpo(-/-)) mice with purified recombinant mouse myeloperoxidase induced a significant antibody response to native myeloperoxidase. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：167 / 178

页数：12

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