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Chemical and structural analysis of an antibody folding intermediate trapped during glycan biosynthesis

Chemical and structural analysis of an antibody folding intermediate trapped during glycan biosynthesis
Chemical and structural analysis of an antibody folding intermediate trapped during glycan biosynthesis

Human IgG Fc glycosylation modulates immunological effector functions such as antibody-dependent cellular cytotoxicity and phagocytosis. Engineering of Fc glycans therefore enables fine-tuning of the therapeutic properties of monoclonal antibodies. The N-linked glycans of Fc are typically complex-type, forming a network of noncovalent interactions along the protein surface of the Cγ2 domain. Here, we manipulate the mammalian glycan-processing pathway to trap IgG1 Fc at sequential stages of maturation, from oligomannose- to hybrid- to complex-type glycans, and show that the Fc is structurally stabilized following the transition of glycans from their hybrid- to complex-type state. X-ray crystallographic analysis of this hybrid-type intermediate reveals that N-linked glycans undergo conformational changes upon maturation, including a flip within the trimannosyl core. Our crystal structure of this intermediate reveals a molecular basis for antibody biogenesis and provides a template for the structure-guided engineering of the protein-glycan interface of therapeutic antibodies.

0002-7863
17554-17563
Bowden, Thomas A.
4b17a588-ac01-4112-807a-8b99a6c20d0f
Baruah, Kavitha
a02f59f3-5e0f-4735-9fdd-639bdbb8f563
Coles, Charlotte H.
8c3ba446-999e-431a-8a4c-aab4ccd3b2dc
Harvey, David J.
8bb24417-3852-4b1f-827b-0d5d2c176744
Yu, Xiaojie
44d52374-eacc-4e23-b7da-c881e6d3a5dd
Song, Byeong Doo
8570f719-c8b1-4772-9e4e-d541f5fcb0a7
Stuart, David I.
2751c230-9d4c-4981-aeb7-cafb51ed749d
Aricescu, A. Radu
8ee3d2bd-7ef7-4e85-862a-e13f48ae0fa6
Scanlan, Christopher N.
04dd1b57-b6fc-414c-8595-08310dbb3d32
Jones, E. Yvonne
6b9004c9-137b-4f43-8a14-c7c83bcaa4be
Crispin, Matthew
cd980957-0943-4b89-b2b2-710f01f33bc9
Bowden, Thomas A.
4b17a588-ac01-4112-807a-8b99a6c20d0f
Baruah, Kavitha
a02f59f3-5e0f-4735-9fdd-639bdbb8f563
Coles, Charlotte H.
8c3ba446-999e-431a-8a4c-aab4ccd3b2dc
Harvey, David J.
8bb24417-3852-4b1f-827b-0d5d2c176744
Yu, Xiaojie
44d52374-eacc-4e23-b7da-c881e6d3a5dd
Song, Byeong Doo
8570f719-c8b1-4772-9e4e-d541f5fcb0a7
Stuart, David I.
2751c230-9d4c-4981-aeb7-cafb51ed749d
Aricescu, A. Radu
8ee3d2bd-7ef7-4e85-862a-e13f48ae0fa6
Scanlan, Christopher N.
04dd1b57-b6fc-414c-8595-08310dbb3d32
Jones, E. Yvonne
6b9004c9-137b-4f43-8a14-c7c83bcaa4be
Crispin, Matthew
cd980957-0943-4b89-b2b2-710f01f33bc9

Bowden, Thomas A., Baruah, Kavitha, Coles, Charlotte H., Harvey, David J., Yu, Xiaojie, Song, Byeong Doo, Stuart, David I., Aricescu, A. Radu, Scanlan, Christopher N., Jones, E. Yvonne and Crispin, Matthew (2012) Chemical and structural analysis of an antibody folding intermediate trapped during glycan biosynthesis. Journal of the American Chemical Society, 134 (42), 17554-17563. (doi:10.1021/ja306068g).

Record type: Article

Abstract

Human IgG Fc glycosylation modulates immunological effector functions such as antibody-dependent cellular cytotoxicity and phagocytosis. Engineering of Fc glycans therefore enables fine-tuning of the therapeutic properties of monoclonal antibodies. The N-linked glycans of Fc are typically complex-type, forming a network of noncovalent interactions along the protein surface of the Cγ2 domain. Here, we manipulate the mammalian glycan-processing pathway to trap IgG1 Fc at sequential stages of maturation, from oligomannose- to hybrid- to complex-type glycans, and show that the Fc is structurally stabilized following the transition of glycans from their hybrid- to complex-type state. X-ray crystallographic analysis of this hybrid-type intermediate reveals that N-linked glycans undergo conformational changes upon maturation, including a flip within the trimannosyl core. Our crystal structure of this intermediate reveals a molecular basis for antibody biogenesis and provides a template for the structure-guided engineering of the protein-glycan interface of therapeutic antibodies.

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Published date: 24 October 2012

Identifiers

Local EPrints ID: 414414
URI: https://eprints.soton.ac.uk/id/eprint/414414
ISSN: 0002-7863
PURE UUID: 350b7d99-7790-4097-a6e1-fac75450de8b
ORCID for Matthew Crispin: ORCID iD orcid.org/0000-0002-1072-2694

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Date deposited: 28 Sep 2017 16:31
Last modified: 22 Oct 2019 00:26

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Contributors

Author: Thomas A. Bowden
Author: Kavitha Baruah
Author: Charlotte H. Coles
Author: David J. Harvey
Author: Xiaojie Yu
Author: Byeong Doo Song
Author: David I. Stuart
Author: A. Radu Aricescu
Author: Christopher N. Scanlan
Author: E. Yvonne Jones
Author: Matthew Crispin ORCID iD

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