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Biotechnol Prog

Title:		Evaluation of immunoglobulins from plant cells
Author(s):		Hein MB; Tang Y; McLeod DA; Janda KD; Hiatt A;
Address:		"Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, California 92037"
Journal Title:		Biotechnol Prog
Year:		1991
Volume:		7
Issue:		5
Page Number:		455 - 461
DOI:		10.1021/bp00011a011
ISSN/ISBN:		8756-7938 (Print) 1520-6033 (Linking)
Abstract:		"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 (Km, Ki, Vmax, and kcat) 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"
Keywords:		"Amino Acid Sequence Animals Antibodies, Monoclonal/genetics/immunology Antigens/immunology Catalysis DNA/genetics *Gene Expression Glycosylation Immunoglobulin gamma-Chains/metabolism Immunoglobulins/*genetics/immunology Mating Factor Mice Molecular Seque;"
Notes:		"MedlineHein, M B Tang, Y McLeod, D A Janda, K D Hiatt, A eng 1991/09/01 Biotechnol Prog. 1991 Sep-Oct; 7(5):455-61. doi: 10.1021/bp00011a011"