EVALUATION OF IMMUNOGLOBULINS FROM PLANT-CELLS

被引：76

作者：

HEIN, MB ^{[1
]}

TANG, Y ^{[1
]}

MCLEOD, DA ^{[1
]}

JANDA, KD ^{[1
]}

HIATT, A ^{[1
]}

机构：

[1] SCRIPPS RES INST, DEPT CHEM, LA JOLLA, CA 92037 USA

来源：

BIOTECHNOLOGY PROGRESS | 1991年 / 7卷 / 05期

关键词：

D O I：

10.1021/bp00011a011

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Expression of cDNA constructs encoding full-length mouse immunoglobulin chains with their native leader sequences or fusion constructs substituting the native leader with a pre-pro sequence derived from Saccharomyces cerevisiae yielded blocked N-termini on the gamma chain or the correct amino terminal sequence on the mature kappa-chain. Lectin binding assays revealed that assembled immunoglobulin complexes contained a glycosylated heavy chain. The attached glycan was resistant to digestion by endoglycosidase H and its lectin binding pattern was distinguishable from that of the mammalian glycan. The results indicated processing of the immunoglobulin carbohydrate in the tobacco Golgi to yield a complex oligosaccharide. Secretion of antibody by protoplasts isolated from regenerated transgenic plants or from suspension callus cells was demonstrated by pulse-chase labeling experiments. When purified, the tobacco-produced antibody was found to possess the antigen binding and catalytic properties of the murine monoclonal antibody. Kinetic parameters (K(m), K(i), V(max, and k(cat) of the tobacco-derived antibody were comparable to those of the mouse-derived antibody. The results in general show that the endomembrane system of tobacco cells possesses cognate mechanisms for the recognition of diverse leader sequences. These signals can be used to initiate the assembly, processing, and secretion by plant cells of complex foreign proteins.

引用

页码：455 / 461

页数：7

共 42 条

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