Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity.
The glycosylation pattern of chCE7, an antineuroblastoma chimeric IgG1, was engineered in Chinese hamster ovary cells with tetracycline-regulated expression of beta(1,4)-N-acetylglucosaminyltransferase III (GnTIII), a glycosyltransferase catalyzing formation of bisected oligosaccharides that have been implicated in antibody-dependent cellular cytotoxicity (ADCC). Measurement of the ADCC activity of chCE7 produced at different tetracycline levels showed an optimal range of GnTIII expression for maximal chCE7 in vitro ADCC activity, and this activity correlated with the level of constant region-associated, bisected complex oligosaccharides determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The new optimized variants of chCE7 exhibit substantial ADCC activity and, hence, may be useful for treatment of neuroblastoma. The strategy presented here should be applicable to optimize the ADCC activity of other therapeutic IgGs.
Institute of Biotechnology, ETH Zürich, Switzerland.
P
P
Umaña
J
J
Jean-Mairet
R
R
Moudry
H
H
Amstutz
J E
JE
Bailey
D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity; D016466:CHO Cells; D002240:Carbohydrate Sequence; D006224:Cricetinae; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics; D009447:Neuroblastoma / Q000276:immunology*; D013752:Tetracycline / Q000494:pharmacology
D007074:Immunoglobulin G; D017351:N-Acetylglucosaminyltransferases; C054599:beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase; D013752:Tetracycline