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Lundblad, Arne
Publications (10 of 17) Show all publications
Liljeblad, M., Lundblad, A. & Påhlsson, P. (2002). Analysis of glycoproteins in cell culture supernatants using a lectin immunosensor technique. Biosensors & bioelectronics, 17(10), 883-891
Open this publication in new window or tab >>Analysis of glycoproteins in cell culture supernatants using a lectin immunosensor technique
2002 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 17, no 10, p. 883-891Article in journal (Refereed) Published
Abstract [en]

A method based on a surface plasmon resonance technique for detection of changes in concentration and glycosylation of proteins in cell culture supernatant is described. The method was used to analyze α1-acid glycoprotein (AGP) produced by a human hepatoma cell line (HepG2). Cell culture supernatant was injected to a BIACORE 2000 instrument and AGP was captured on the sensor chip by immobilized antibodies. The captured glycoprotein was then analyzed for content of carbohydrate epitopes using three different lectins, Aleuria aurantia lectin (AAL), Sambucus nigra agglutinin (SNA), and Triticum vulgaris agglutinin (wheat germ agglutinin, WGA). The method was used to analyze changes in concentration and glycosylation of AGP produced by HepG2 cells grown with or without three different cytokines, interleukin-1β (IL-1β), interleukin-6 (IL-6), and transforming growth factor β-1 (TGFβ1). Using the described method it was shown that when HepG2 cells were grown in the presence of IL-6 both AGP concentration and fucosylation increased. When HepG2 cells instead were grown in the presence of TGFβ1 AGP fucosylation increased whereas AGP concentration decreased.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25226 (URN)10.1016/S0956-5663(02)00111-2 (DOI)9665 (Local ID)9665 (Archive number)9665 (OAI)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
Rydén, I., Påhlsson, P., Lundblad, A. & Skogh, T. (2002). Fucosylation of α1-acid glycoprotein (orosomucoid) compared with traditional biochemical markers of inflammation in recent onset rheumatoid arthritis. Clinica Chimica Acta, 317(1-2), 221-229
Open this publication in new window or tab >>Fucosylation of α1-acid glycoprotein (orosomucoid) compared with traditional biochemical markers of inflammation in recent onset rheumatoid arthritis
2002 (English)In: Clinica Chimica Acta, ISSN 0009-8981, E-ISSN 1873-3492, Vol. 317, no 1-2, p. 221-229Article in journal (Refereed) Published
Abstract [en]

Background: Fucosylation of α1-acid glycoprotein (AGP, orosomucoid) has previously been found to be increased in patients with rheumatoid arthritis. Furthermore, the degree of fucosylation has been suggested to reflect disease activity. Therefore, we investigated the fucosylation of AGP in 131 patients (96 women and 35 men) with recent onset rheumatoid arthritis (RA). We compared the results with traditional biochemical markers of inflammation, i.e. plasma concentrations of AGP (P-AGP), and C-reactive protein (P-CRP).

Methods: AGP fucosylation measured with a novel lectin enzyme-linked immunosorbent assay (ELISA) was compared with a disease activity score (DAS28) and its components, and with P-AGP, and P-CRP at the time of diagnosis, and at a follow-up visit 1 year later.

Results: Both men and women with RA had increased AGP fucosylation compared to healthy individuals. We found a weak correlation between AGP fucosylation and DAS28 only in men. In men with initially increased AGP fucosylation, the level of fucosylation correlated with the change in DAS28 during the first year following diagnosis.

Conclusion: We conclude that AGP fucosylation is not superior to traditional markers of disease activity in RA. However, AGP fucosylation may give some additional information to traditional biochemical markers on the disease progression in men.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25259 (URN)10.1016/S0009-8981(01)00803-8 (DOI)9699 (Local ID)9699 (Archive number)9699 (OAI)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
Bengtson, P., Lundblad, A., Larson, G. & Påhlsson, P. (2002). Polymorphonuclear Leukocytes from Individuals Carrying the G329A Mutation in the α1,3-Fucosyltransferase VII Gene (FUT7) Roll on E- and P-Selectins. Journal of Immunology, 169(7), 3940-3946
Open this publication in new window or tab >>Polymorphonuclear Leukocytes from Individuals Carrying the G329A Mutation in the α1,3-Fucosyltransferase VII Gene (FUT7) Roll on E- and P-Selectins
2002 (English)In: Journal of Immunology, ISSN 0022-1767, E-ISSN 1550-6606, Vol. 169, no 7, p. 3940-3946Article in journal (Refereed) Published
Abstract [en]

We recently identified several individuals carrying a missense mutation (G329A; Arg110-Gln) in the FUT7 gene encoding fucosyltransferase VII. This enzyme is involved in the biosynthesis of the sialyl Lewis x (Lex) epitope on human leukocytes, which has been identified as an important component of leukocyte ligands for E- and P-selectin. No enzyme activity was measurable in expression studies in COS-7 cells using the mutated FUT7 construct. One of the identified individuals carried this mutation homozygously. Flow cytometry analysis of polymorphonuclear leukocytes (PMN) from this individual showed a nearly complete absence of staining with mAbs directed against sialyl Lex and a diminished staining with an E-selectin IgG chimera. However, staining with P-selectin IgG chimera and Abs directed against P-selectin glycoprotein ligand-1 was not affected by the mutation. PMN from the homozygously mutated individual was further analyzed in an in vitro flow chamber assay. The number of rolling PMN and the rolling velocities on both E- and P-selectin were in the range of PMN from nonmutated individuals. FUT4 and FUT7 mRNA was quantified in PMN isolated from individuals carrying the FUT7 mutation. It was found that PMN from both FUT7 homozygously and heterozygously mutated individuals exhibited an elevated expression of FUT4 mRNA compared with PMN from FUT7 nonmutated individuals. The elevated expression of fucosyltransferase IV was reflected as an increased expression of the Lex and CD65s Ags on PMN from these individuals. The significance of the mutation was supported by transfection of BJAB cells.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-24924 (URN)9329 (Local ID)9329 (Archive number)9329 (OAI)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
Liljeblad, M., Rydén, I., Ohlson, S., Lundblad, A. & Påhlsson, P. (2001). A Lectin Immunosensor Technique for Determination of α1-Acid Glycoprotein Fucosylation. Analytical Biochemistry, 288(2), 216-224
Open this publication in new window or tab >>A Lectin Immunosensor Technique for Determination of α1-Acid Glycoprotein Fucosylation
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2001 (English)In: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 288, no 2, p. 216-224Article in journal (Refereed) Published
Abstract [en]

The fucosylation of α1-acid glycoprotein (AGP), an acute-phase protein, is known to change in association with inflammatory diseases. Thus, fucosylation of AGP could be a potential diagnostic or prognostic marker. The change in fucosylation has previously been investigated using crossed affinoimmunoelectrophoresis, high-pH anion-exchange chromatography, and lectin ELISA. This study describes a surface plasmon resonance-based affinity biosensor assay for quantification of the fucosylation of AGP. Diluted EDTA plasma or serum was injected directly in a BIACORE 2000 biosensor. AGP was captured on the sensor surface using immobilized antibodies and a fucose-binding lectin from Aleuria aurentia was then used for the detection of fucosylation. The feature of the biosensor makes it possible to determine both the amount of bound AGP and the amount of bound lectin. Using a calibration curve it was possible to obtain a fucosylation ratio that was independent of AGP concentration. The assay was validated against a lectin ELISA and used to follow inflammation in patients with severe burns.

Keywords
a1-acid glycoprotein, Aleuria aurentia lectin, Fucose, Glycosylation, Surface plasmon resonance
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25224 (URN)10.1006/abio.2000.4905 (DOI)9663 (Local ID)9663 (Archive number)9663 (OAI)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
Bengtson, P., Larson, C., Lundblad, A., Larson, G. & Påhlsson, P. (2001). Identification of a Missense Mutation (G329A; Arg110→ Gln) in the Human FUT7 Gene. Journal of Biological Chemistry, 276(34), 31575-31582
Open this publication in new window or tab >>Identification of a Missense Mutation (G329A; Arg110→ Gln) in the Human FUT7 Gene
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2001 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 276, no 34, p. 31575-31582Article in journal (Refereed) Published
Abstract [en]

The human FUT7 gene codes for the α1,3-fucosyltransferase VII (Fuc-TVII), which is involved in the biosynthesis of the sialyl Lewis x (SLex) epitope on human leukocytes. The FUT7 gene has so far been considered to be monomorphic. Neutrophils isolated from patients with ulcerative colitis were examined for apparent alterations in protein glycosylation patterns by Western blot analysis using monoclonal antibodies directed against SLex and SLex-related epitopes. One individual showed lower levels of SLex expression and an elevated expression of CD65s compared to controls. The coding regions of the FUT7 gene from this individual were cloned, and a G329A point mutation (Arg110 → Gln) was found in one allele, whereas the other FUT7 allele was wild type. No Fuc-TVII enzyme activity was detected in COS-7 cells transiently transfected with the mutated FUT7 construct. TheFUT7 Arg110 is conserved in all previously cloned vertebrate α1,3-fucosyltransferases. Polymerase chain reaction followed by restriction enzyme cleavage was used to screen 364 unselected Caucasians for the G329A mutation, and a frequency of ≤1% for this mutation was found (3 heterozygotes). Genetic characterization of the family members of one of the additional heterozygotes identified one individual carrying the G329A mutation in both FUT7alleles. Peripheral blood neutrophils of this homozygously mutated individual showed a lowered expression of SLex and an elevated expression of CD65s when analyzed by Western blot and flow cytometry. The homozygous individual was diagnosed with ulcer disease, non-insulin-dependent diabetes, osteoporosis, spondyloarthrosis, and Sjögren's syndrome but had no history of recurrent bacterial infections or leukocytosis.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-47286 (URN)10.1074/jbc.M104165200 (DOI)
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2017-12-13Bibliographically approved
Liljeblad, M., Lundblad, A. & Påhlsson, P. (2000). Analysis of agalacto-IgG in rheumatoid arthritis using surface plasmon resonance. Glycoconjugate Journal, 17(5), 323-329
Open this publication in new window or tab >>Analysis of agalacto-IgG in rheumatoid arthritis using surface plasmon resonance
2000 (English)In: Glycoconjugate Journal, ISSN 0282-0080, E-ISSN 1573-4986, Vol. 17, no 5, p. 323-329Article in journal (Refereed) Published
Abstract [en]

It is well established that IgG from rheumatoid arthritis (RA) patients are less galactosylated than IgG from normal individuals. Determination of agalacto-IgG may therefore aid in diagnosis and treatment of RA. The decrease in galactosylation of IgG leads to an increase in terminal N-acetylglucosamine residues, which can be detected using a specific lectin from Psathyrella velutina. In the present study IgG from RA and control serum was purified using affinity chromatography. The samples were then, after reduction, analyzed on a BIOCORE® 2000 system with immobilized Psathyrella velutina lectin. Using this technique it was possible to discriminate between IgG from RA patients and IgG from control individuals with respect to its content of IgG with terminal N-acetylglucosamine. The affinity biosensor technique makes it possible to detect binding without labeling or using secondary antibodies.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25225 (URN)10.1023/A:1007169621518 (DOI)9664 (Local ID)9664 (Archive number)9664 (OAI)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
Landberg, E., Huang, Y., Strömqvist, M., Mechref, Y., Hansson, L., Lundblad, A., . . . Påhlsson, P. (2000). Changes in Glycosylation of Human Bile-Salt-Stimulated Lipase during Lactation. Archives of Biochemistry and Biophysics, 377(2), 246-254
Open this publication in new window or tab >>Changes in Glycosylation of Human Bile-Salt-Stimulated Lipase during Lactation
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2000 (English)In: Archives of Biochemistry and Biophysics, ISSN 0003-9861, E-ISSN 1096-0384, Vol. 377, no 2, p. 246-254Article in journal (Refereed) Published
Abstract [en]

Bile-salt-stimulated lipase (BSSL) is an enzyme in human milk, which is important for the fat digestion in the newborn infant. BSSL is highly glycosylated and includes one site for N-glycosylation and several sites for O-glycosylation. BSSL has previously been found to express Lewis a, Lewis b, and Lewis x carbohydrate antigens. In this study, glycosylation of BSSL was studied at different times during lactation. BSSL was purified from milk collected individually from four donors at several different times during the first 6 months of lactation. The BSSL glycans were characterized through monosaccharide analysis, high-pH anion-exchange chromatography, matrix-assisted laser desorption–ionization mass spectrometry, and ELISA. Both total carbohydrate content and relative amount of sialic acid were higher in BSSL from the first lactation month as compared to BSSL from milk collected later in lactation. BSSL from the first lactation month also showed a different composition of sialylated O-linked glycans and the N-linked oligosaccharides consisted of lower amounts of fucosylated structures compared to later in lactation. We also found a gradual increase in the expression of the carbohydrate epitope Lewis x on BSSL throughout the lactation period. This study shows that glycosylation of BSSL is dependent on blood group phenotype of the donor and changes substantially during the lactation period.

Keywords
bile-salt-stimulated lipase, glycosylation, Lewis x, human milk
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25198 (URN)10.1006/abbi.2000.1778 (DOI)9637 (Local ID)9637 (Archive number)9637 (OAI)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
Rydén, I., Lundblad, A. & Påhlsson, P. (1999). Lectin ELISA for Analysis of α1-Acid Glycoprotein Fucosylation in the Acute Phase Response. Clinical Chemistry, 45(11), 2010-2012
Open this publication in new window or tab >>Lectin ELISA for Analysis of α1-Acid Glycoprotein Fucosylation in the Acute Phase Response
1999 (English)In: Clinical Chemistry, ISSN 0009-9147, E-ISSN 1530-8561, Vol. 45, no 11, p. 2010-2012Article in journal (Refereed) Published
Abstract [en]

No abtract available.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25261 (URN)9701 (Local ID)9701 (Archive number)9701 (OAI)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13Bibliographically approved
Lindberg, G., Råstam, L., Nilsson-Ehle, P., Lundblad, A., Ranstam, J., Folsom, A. & Burke, G. (1999). Serum sialic acid and sialoglycoproteins in asymptomatic carotid artery atherosclerosis.. Atherosclerosis, 146, 65-69
Open this publication in new window or tab >>Serum sialic acid and sialoglycoproteins in asymptomatic carotid artery atherosclerosis.
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1999 (English)In: Atherosclerosis, ISSN 0021-9150, E-ISSN 1879-1484, Vol. 146, p. 65-69Article in journal (Refereed) Published
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-25260 (URN)9700 (Local ID)9700 (Archive number)9700 (OAI)
Available from: 2009-10-07 Created: 2009-10-07 Last updated: 2017-12-13
Liljeblad, M., Lundblad, A., Ohlson, S. & Påhlsson, P. (1998). Detection of low-molecular-weight heparin oligosaccharides (Fragmin™) using surface plasmon resonance. Journal of Molecular Recognition, 11(1-6), 191-193
Open this publication in new window or tab >>Detection of low-molecular-weight heparin oligosaccharides (Fragmin™) using surface plasmon resonance
1998 (English)In: Journal of Molecular Recognition, ISSN 0952-3499, E-ISSN 1099-1352, Vol. 11, no 1-6, p. 191-193Article in journal (Refereed) Published
Abstract [en]

During the last decades there has been a growing realization of the central biological role that oligosaccharides and oligosaccharide–protein interactions play. One of the most striking examples is the use of heparin and low-molecular-weight heparin oligosaccharides (Fragmin™) to modify blood coagulation. Several monoclonal antibodies directed against glycosaminoglycan structures have been produced. However, their clinical use is limited by the difficulty of detection systems for oligosaccharides. In the present study we used a monoclonal antibody directed against heparin oligosaccharides prepared by partial nitrous acid deamination of heparin. Using a biosensor (BIAcore™), purified antibody was immobilized on sensor surfaces and binding of oligosaccharide was measured by surface plasmon resonance. Using this technique, it was possible to quantitate low-molecular-weight heparin oligosaccharides in nanomolar concentrations.

Place, publisher, year, edition, pages
John Wiley and Sons, 1998
Keywords
surface plasmon resonance, low-molecular-weight heparin, oligosaccharides
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-75800 (URN)10.1002/(SICI)1099-1352(199812)11:1/6<191::AID-JMR421>3.0.CO;2-4 (DOI)000078789000043 ()
Available from: 2012-03-12 Created: 2012-03-12 Last updated: 2017-12-07Bibliographically approved
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