| 1. |
- Rich, Rebecca L., et al.
(författare)
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A global benchmark study using affinity-based biosensors
- 2009
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Ingår i: Analytical Biochemistry. - : Elsevier BV. - 0003-2697 .- 1096-0309. ; 386:2, s. 194-216
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Tidskriftsartikel (refereegranskat)abstract
- To explore the variability in biosensor studies, 150 participants from 20 countries were given the same protein samples and asked to determine kinetic rate constants for the interaction. We chose a protein system that was amenable to analysis using different biosensor platforms as well as by users of different expertise levels. The two proteins (a 50-kDa Fab and a 60-kDa glutathione S-transferase [GST] antigen) form a relatively high-affinity complex, so participants needed to optimize several experimental parameters, including ligand immobilization and regeneration conditions as well as analyte concentrations and injection/dissociation times. Although most participants collected binding responses that could be fit to yield kinetic parameters, the quality of a few data sets could have been improved by optimizing the assay design. Once these outliers were removed, the average reported affinity across the remaining panel of participants was 620 pM with a standard deviation of 980 pM. These results demonstrate that when this biosensor assay was designed and executed appropriately, the reported rate constants were consistent, and independent of which protein was immobilized and which biosensor was used.
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| 2. |
- Balogh, Péter, 1982-
(författare)
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Perpetual borders : German-Polish cross-border contacts in the Szczecin area
- 2014
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Borderlands are often peripheral geographically, administratively, and economically. A particularly illustrative case is the Szczecin area at the border between Poland and Germany, where a large city on one side neighbours to a sparsely populated hinterland on the other. There is a number of similar cases throughout Europe, but studies on them point to a mixed level of linkages following the opening and removal of the physical border.At the project’s start there were few if any studies on the Szczecin area per se, which was here studied through various methods. On the one hand, different pre-EU enlargement plans and visions for the area’s development were compared with practices and realities of recent years. This shows that earlier imaginations on the development potentials have not quite materialised, although some of them were probably too optimistic and ambitious from the beginning. Some of the area’s potentials following EU-enlargement have been more successfully exploited than others, and disproportionately by actors coming from outside. On the other hand, cross-border contacts were studied in the discourses on and attitudes towards the other side among local and regional elites, and among local residents more generally. This revealed a polarised attitudinal landscape, not least when compared to country-wide opinion surveys in both Germany and Poland. This is in line with other studies showing that identities are particularly accentuated in border situations, where the Other is more frequently encountered.These results support recent investigations pointing to a continued relevance of the border even after the physical barriers are removed. At the same time, another contribution of this work to border studies is that the time and contingency of the importance of identities and of the border needs more attention. In the Szczecin area, awareness of national identities and of the boundary appeared to be particularly high just after changes in the border’s status occurred – i.e. in 1989–1991, and then around the years 2007–2010. But while its importance may be fluctuating over time, given the opportunities and resources the boundary provides it will always be maintained in some forms.
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| 3. |
- Dzieciatkowska, Monika, et al.
(författare)
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Rapid method for sensitive screening of oligosaccharide epitopes in the lipooligosaccharide from Campylobacter jejuni strains isolated from Guillain-Barre syndrome and Miller Fisher syndrome patients
- 2008
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Ingår i: Journal of Clinical Microbiology. - 0095-1137 .- 1098-660X. ; 46:10, s. 3429-3436
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Tidskriftsartikel (refereegranskat)abstract
- Campylobacter jejuni lipooligosaccharide (LOS) can trigger Guillain-Barre syndrome (GBS) due to its similarity to human gangliosides. Rapid and accurate structural elucidation of the LOS glycan of a strain isolated from a GBS patient could help physicians determine the spectrum of anti-ganglioside antibodies likely to be found and therefore provide valuable assistance in establishing an appropriate course of treatment. The ability of implemented mass spectrometry-based approaches in a clinical setting has been limited by the laborious and time-consuming nature of the protocols, typically 3 to 4 days, used to prepare LOS. In order to improve the analytical throughput, microwave-assisted enzymatic digestion was investigated. In this study, the bacterial cells were suspended in 50 mu l of 20 mM ammonium acetate buffer containing DNase and RNase and treated by direct microwave irradiation for 3 min. Then, proteinase K was added and the samples were again microwaved. The intact LOS samples were analyzed using electrophoresis-assisted open-tubular liquid chromatography-mass spectrometry. The reliability of the rapid, high-throughput technique was demonstrated through analysis of LOS glycans from 73 C. jejuni strains. The structure was elucidated using material from a single colony. The total time for sample preparation and MS analysis is less than 60 min.
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| 4. |
- Houliston, R. Scott, et al.
(författare)
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A Haemophilus influenzae strain associated with fisher syndrome expresses a novel disialylated ganglioside mimic
- 2007
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Ingår i: Biochemistry. - : American Chemical Society (ACS). - 0006-2960 .- 1520-4995. ; 46:27, s. 8164-8171
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Tidskriftsartikel (refereegranskat)abstract
- The non-typeable Haemophilus influenzae strain DH1 was isolated from a 25 year old male patient with Fisher syndrome, a postinfectious autoimmune condition characterized by the presence of anti-GQ1b IgG antibodies that target and initiate damage to peripheral nerves. DH1 was found to display an alpha NeuAc(2-8)alpha NeuAc(2-3)beta Gal branch bound to the tetraheptosyl backbone core of its lipooligosaccharide (LOS). The novel sialylation pattern was found to be dependent on the activity of a bifunctional sialyltransferase, Lic3B, which catalyzes the addition of both the terminal and subterminal sialic acid residues. Patient serum IgGs bind to DH1 LOS, and the reactivity is significantly influenced by the presence of sialylated glycoforms. The display by DH1, of a surface glycan that mimics the terminal trisaccharide portion of disialosyl-containing gangliosides, provides strong evidence for its involvement in the development of Fisher syndrome.
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