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Pichia pastoris-pro...
Pichia pastoris-produced mucin-type fusion proteins with multivalent O-glycan substitution as targeting molecules for mannose-specific receptors of the immune system
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- Gustafsson, Anki (author)
- Recopharma AB, Huddinge
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- Sjöblom, Magnus (author)
- Luleå tekniska universitet,Industriell miljö- och processteknik
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- Strindelius, Lena (author)
- Recopharma AB, Huddinge
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- Johansson, Thomas (author)
- Recopharma AB, Huddinge
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- Fleckenstein, Tilly (author)
- Recopharma AB, Huddinge
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- Chatzissavidou, Nathalie (author)
- Recopharma AB, Huddinge
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- Lindberg, Linda (author)
- Absorber AB, Stockholm
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- Ångström, Jonas, 1950 (author)
- Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi,Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
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- Rova, Ulrika (author)
- Luleå tekniska universitet,Industriell miljö- och processteknik
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- Holgersson, Jan (author)
- Karolinska Institutet
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(creator_code:org_t)
- 2011-04-07
- 2011
- English.
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In: Glycobiology. - : Oxford University Press (OUP). - 0959-6658 .- 1460-2423. ; 21:8, s. 1071-1086
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Subject headings
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- Mannose-binding proteins like the macrophage mannose receptor (MR), the dendritic cell-specific intercellular adhesion molecule-3 grabbing non-integrin (DC-SIGN) and mannose-binding lectin (MBL) play crucial roles in both innate and adaptive immune responses. Immunoglobulin fusion proteins of the P-selectin glycoprotein ligand-1 (PSGL-1/mIgG2b) carrying mostly O-glycans and, as a control, the a1-acid glycoprotein (AGP/mIgG2b) carrying mainly N-linked glycans were stably expressed in the yeast Pichia pastoris. P. pastoris-produced PSGL-1/mIgG2b was shown to carry O-glycans that mediated strong binding to mannose-specific lectins in a lectin array and were susceptible to cleavage by a-mannosidases including an a1,2- but not an a1,6-mannosidase. Electrospray ionization - ion trap mass spectrometry (ESI-MS) confirmed the presence of O-glycans containing up to 9 hexoses with the penta- and hexasaccharides being the predominant ones. a1,2- and a1,3-linked, but not a1,6-linked, mannose residues were detected by 1H-nuclear magnetic resonance (1H-NMR) spectroscopy confirming the results of the mannosidase cleavage. The apparent equilibrium dissociation constants for binding of PNGase F-treated mannosylated PSGL-1/mIgG2b to MR, DC-SIGN and MBL were shown by surface plasmon resonance to be 126, 56 and 16 nM, respectively. In conclusion, PSGL-1/mIgG2b expressed in P. pastoris carried O-glycans mainly comprised of a-linked mannoses and with up to nine residues. It bound mannose-specific receptors with high apparent affinity and may become a potent targeting molecule for these receptors in vivo.
Subject headings
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik -- Bioprocessteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology -- Bioprocess Technology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Keyword
- Biokemisk processteknik
- Biochemical Process Engineering
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Gustafsson, Anki
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Sjöblom, Magnus
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Strindelius, Len ...
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Johansson, Thoma ...
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Fleckenstein, Ti ...
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Chatzissavidou, ...
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show more...
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Lindberg, Linda
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Ångström, Jonas, ...
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Rova, Ulrika
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Holgersson, Jan
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- About the subject
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- ENGINEERING AND TECHNOLOGY
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ENGINEERING AND ...
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and Industrial Biote ...
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and Bioprocess Techn ...
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Medical Biotechn ...
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and Medical Biotechn ...
- Articles in the publication
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Glycobiology
- By the university
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Luleå University of Technology
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University of Gothenburg
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Karolinska Institutet