Journal article
Binding of DC-SIGN to glycoproteins expressed in glycoengineered Pichia pastoris
Journal of immunological methods, v 386(01-02), pp 34-42
14 Dec 2012
PMID: 22982058
Abstract
Previous studies have shown that glycoproteins expressed in wild-type Pichia pastoris bind to Dendritic cell-SIGN (DC-Specific Intercellular adhesion molecule-3 Grabbing Nonintegrin), a mannose-binding receptor found on dendritic cells in peripheral tissues which is involved in antigen presentation and the initiation of an immune response. However, the binding of DC-SIGN to glycoproteins purified from P. pastoris strains engineered to express humanized N- and O-linked glycans has not been tested to date. In this study, the binding of glycoproteins with specific high-mannose or human N- and O-linked glycan structures to DC-SIGN was tested. Proteins with humanized N-glycans including Man5 structures and O-glycans (up to as many as 24) with single mannose chain length showed DC-SIGN binding that was comparable to that measured for a CHO-produced IgG1 which lacks O-linked mannose. Glycoproteins with wild-type N-glycans and mannotriose and higher O-glycans bound to DC-SIGN in a manner that was strongly inhibited by either the use of enzymatic N-deglycosylation or sodium meta-periodate oxidation. Mannan purified from humanized P. pastoris also showed lower ability to inhibit DC-SIGN binding to glycoproteins with wild type fungal glycosylation than mannan purified from wild type strains. This study shows that humanized P. pastoris can produce glycoproteins that do not bind to DC-SIGN.
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Details
- Title
- Binding of DC-SIGN to glycoproteins expressed in glycoengineered Pichia pastoris
- Creators
- Michael C. Cukan - Merck & Co., Inc., Rahway, NJ, USA (United States)Daniel Hopkins - Merck & Co., Inc., Rahway, NJ, USA (United States)Irina Burnina - Merck & Co., Inc., Rahway, NJ, USA (United States)Michelle Button - Merck & Co., Inc., Rahway, NJ, USA (United States)Erin Giaccone - Merck & Co., Inc., Rahway, NJ, USA (United States)Nga Rewa Houston-Cummings - Merck & Co., Inc., Rahway, NJ, USA (United States)Youwei Jiang - Purification Development, GlycoFi, Inc., Merck & Co., 21 Lafayette Street, Suite 200, Lebanon, New Hampshire 03766, United StatesFang Li - Merck & Co., Inc., Rahway, NJ, USA (United States)Muralidhar Mallem - Merck & Co., Inc., Rahway, NJ, USA (United States)Teresa Mitchell - Merck & Co., Inc., Rahway, NJ, USA (United States)Renée Moore - Merck & Co., Inc., Rahway, NJ, USA (United States)Adam Nylen - Merck & Co., Inc., Rahway, NJ, USA (United States)Bianka Prinz - Merck & Co., Inc., Rahway, NJ, USA (United States)Sandra Rios - Merck & Co., Inc., Rahway, NJ, USA (United States)Nathan Sharkey - Merck & Co., Inc., Rahway, NJ, USA (United States)Dongxing Zha - Merck & Co., Inc., Rahway, NJ, USA (United States)Stephen Hamilton - Merck & Co., Inc., Rahway, NJ, USA (United States)Huijuan Li - Merck & Co., Inc., Rahway, NJ, USA (United States)Terrance A. Stadheim - Merck & Co., Inc., Rahway, NJ, USA (United States)
- Publication Details
- Journal of immunological methods, v 386(01-02), pp 34-42
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Epidemiology and Biostatistics
- Web of Science ID
- WOS:000311132800005
- Scopus ID
- 2-s2.0-84867482896
- Other Identifier
- 991021448055504721
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemical Research Methods
- Immunology