Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system

P.H.C. van Berkel; J. Gerritsen; E. van Voskuilen; G. Perdok; T. Vink; J.G.J. van de Winkel; P.W.H.I. Parren

doi:10.1002/bit.22535

Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system

P.H.C. van Berkel, J. Gerritsen, E. van Voskuilen, G. Perdok, T. Vink, J.G.J. van de Winkel, P.W.H.I. Parren

Center for Translational Immunology (CTI)

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Rapid production of recombinant human IgG with improved antibody dependent cell-mediated cytotoxicity (ADCC) effector function is presented. The technique employs transient expression of IgG in suspension growing HEK-293F cells in the presence of the glycosidase inhibitor kifunensine. The procedure takes ∼7 days, provided that expression plasmids encoding the IgG of interest are available. Kifunensine inhibits the N-linked glycosylation pathway of HEK-293F cells in the endoplasmatic reticulum, resulting in IgG with oligomannose type glycans lacking core-fucose. IgG1 transiently produced in kifunensine- treated HEK-293F cells has improved affinity for the FcγRIIIA molecule as measured in an ELISA based assay, and almost eightfold enhanced ADCC using primary peripheral blood mononuclear effector cells. Biotechnol. Bioeng. 2010; 105: 350–357. © 2009 Wiley Periodicals, Inc.

Original language	English
Pages (from-to)	350-357
Number of pages	8
Journal	Biotechnology and Bioengineering
Volume	105
Issue number	2
DOIs	https://doi.org/10.1002/bit.22535
Publication status	Published - 2010

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title = "Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system",

abstract = "Rapid production of recombinant human IgG with improved antibody dependent cell-mediated cytotoxicity (ADCC) effector function is presented. The technique employs transient expression of IgG in suspension growing HEK-293F cells in the presence of the glycosidase inhibitor kifunensine. The procedure takes ∼7 days, provided that expression plasmids encoding the IgG of interest are available. Kifunensine inhibits the N-linked glycosylation pathway of HEK-293F cells in the endoplasmatic reticulum, resulting in IgG with oligomannose type glycans lacking core-fucose. IgG1 transiently produced in kifunensine- treated HEK-293F cells has improved affinity for the FcγRIIIA molecule as measured in an ELISA based assay, and almost eightfold enhanced ADCC using primary peripheral blood mononuclear effector cells. Biotechnol. Bioeng. 2010; 105: 350–357. {\textcopyright} 2009 Wiley Periodicals, Inc.",

author = "{van Berkel}, P.H.C. and J. Gerritsen and {van Voskuilen}, E. and G. Perdok and T. Vink and {van de Winkel}, J.G.J. and P.W.H.I. Parren",

year = "2010",

doi = "10.1002/bit.22535",

language = "English",

volume = "105",

pages = "350--357",

journal = "Biotechnology and Bioengineering",

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TY - JOUR

T1 - Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system

AU - van Berkel, P.H.C.

AU - Gerritsen, J.

AU - van Voskuilen, E.

AU - Perdok, G.

AU - Vink, T.

AU - van de Winkel, J.G.J.

AU - Parren, P.W.H.I.

PY - 2010

Y1 - 2010

N2 - Rapid production of recombinant human IgG with improved antibody dependent cell-mediated cytotoxicity (ADCC) effector function is presented. The technique employs transient expression of IgG in suspension growing HEK-293F cells in the presence of the glycosidase inhibitor kifunensine. The procedure takes ∼7 days, provided that expression plasmids encoding the IgG of interest are available. Kifunensine inhibits the N-linked glycosylation pathway of HEK-293F cells in the endoplasmatic reticulum, resulting in IgG with oligomannose type glycans lacking core-fucose. IgG1 transiently produced in kifunensine- treated HEK-293F cells has improved affinity for the FcγRIIIA molecule as measured in an ELISA based assay, and almost eightfold enhanced ADCC using primary peripheral blood mononuclear effector cells. Biotechnol. Bioeng. 2010; 105: 350–357. © 2009 Wiley Periodicals, Inc.

AB - Rapid production of recombinant human IgG with improved antibody dependent cell-mediated cytotoxicity (ADCC) effector function is presented. The technique employs transient expression of IgG in suspension growing HEK-293F cells in the presence of the glycosidase inhibitor kifunensine. The procedure takes ∼7 days, provided that expression plasmids encoding the IgG of interest are available. Kifunensine inhibits the N-linked glycosylation pathway of HEK-293F cells in the endoplasmatic reticulum, resulting in IgG with oligomannose type glycans lacking core-fucose. IgG1 transiently produced in kifunensine- treated HEK-293F cells has improved affinity for the FcγRIIIA molecule as measured in an ELISA based assay, and almost eightfold enhanced ADCC using primary peripheral blood mononuclear effector cells. Biotechnol. Bioeng. 2010; 105: 350–357. © 2009 Wiley Periodicals, Inc.

U2 - 10.1002/bit.22535

DO - 10.1002/bit.22535

M3 - Article

SN - 0006-3592

VL - 105

SP - 350

EP - 357

JO - Biotechnology and Bioengineering

JF - Biotechnology and Bioengineering

IS - 2

ER -

Rapid production of recombinant human IgG With improved ADCC effector function in a transient expression system

Abstract

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