Engineering Bispecificity into a Single Albumin-Binding Domain

Bispecific antibodies as well as non-immunoglobulin based bispecific affinity proteins are considered to have a very high potential in future biotherapeutic applications. In this study, we report on a novel approach for generation of extremely small bispecific proteins comprised of only a single structural domain. Binding to tumor necrosis factor-α (TNF-α) was engineered into an albumin-binding domain while still retaining the original affinity for albumin, resulting in a bispecific protein composed of merely 46 amino acids. By diversification of the non albumin-binding side of the three-helix bundle domain, followed by display of the resulting library on phage particles, bispecific single-domain proteins were isolated using selections with TNF-α as target. Moreover, based on the obtained sequences from the phage selection, a second-generation library was designed in order to further increase the affinity of the bispecific candidates. Staphylococcal surface display was employed for the affinity maturation, enabling efficient isolation of improved binders as well as multiparameter-based sortings with both TNF-α and albumin as targets in the same selection cycle. Isolated variants were sequenced and the binding to albumin and TNF-α was analyzed. This analysis revealed an affinity for TNF-α below 5 nM for the strongest binders. From the multiparameter sorting that simultaneously targeted TNF-α and albumin, several bispecific candidates were isolated with high affinity to both antigens, suggesting that cell display in combination with fluorescence activated cell sorting is a suitable technology for engineering of bispecificity. To our knowledge, the new binders represent the smallest engineered bispecific proteins reported so far. Possibilities and challenges as well as potential future applications of this novel strategy are discussed

Praksisstudienes omfang og organisering i bioingeniørutdanningen

Source at https://www.bioingenioren.no/arkiv/.Siden 2005 har omfang og innhold i bioingeniørutdanningen vært definert av en nasjonal rammeplan for å sikre likeverdig sluttkompetanse for alle kandidater. Fra 2019 ble en ny forskrift om nasjonale retningslinjer fastsatt. Både tidligere rammeplan og nye retningslinjer har regulert læringsutbyttene og omfang av praksis i utdanningen. I denne studien kartlegges omfang og organisering av praksisstudiene ved de ulike studieprogrammene, dette for å undersøke forskjeller og likheter og tilrettelegge for at studiestedene kan lære av hverandre. Det er innhentet data fra hvert studieprogram for omfang og organisering av veiledet intern praksis, ekstern praksis samt bacheloroppgaven. Dette ble kartlagt for alle studiestedene før og etter innføring av nye retningslinjer. Resultatene viser ulikheter i omfang og organisering av praksisstudiene og bacheloroppgaven mellom studieprogrammene i Norge og da særlig i organisering av intern praksis. De nye retningslinjene åpner samtidig opp for en videre fortolkning av praksisbegrepet og av krav til omfang og organisering av praksis. For å sikre likeverdig sluttkompetanse for ferdigutdannede, uavhengig av studiested, er det behov for i større grad enes om hva som gir tilstrekkelig omfang og god kvalitet i praksis i bioingeniørutdanningen. Denne deskriptive studien viser behov for videre forskningssamarbeid mellom studiestedene, for å utvikle en bioingeniørutdanning av best mulig kvalitet for fremtiden

Praksisstudienes omfang og organisering i bioingeniørutdanningen

Siden 2005 har omfang og innhold i bioingeniørutdanningen vært definert av en nasjonal rammeplan for å sikre likeverdig sluttkompetanse for alle kandidater. Fra 2019 ble en ny forskrift om nasjonale retningslinjer fastsatt. Både tidligere rammeplan og nye retningslinjer har regulert læringsutbyttene og omfang av praksis i utdanningen. I denne studien kartlegges omfang og organisering av praksisstudiene ved de ulike studieprogrammene, dette for å undersøke forskjeller og likheter og tilrettelegge for at studiestedene kan lære av hverandre. Det er innhentet data fra hvert studieprogram for omfang og organisering av veiledet intern praksis, ekstern praksis samt bacheloroppgaven. Dette ble kartlagt for alle studiestedene før og etter innføring av nye retningslinjer. Resultatene viser ulikheter i omfang og organisering av praksisstudiene og bacheloroppgaven mellom studieprogrammene i Norge og da særlig i organisering av intern praksis. De nye retningslinjene åpner samtidig opp for en videre fortolkning av praksisbegrepet og av krav til omfang og organisering av praksis. For å sikre likeverdig sluttkompetanse for ferdigutdannede, uavhengig av studiested, er det behov for i større grad enes om hva som gir tilstrekkelig omfang og god kvalitet i praksis i bioingeniørutdanningen. Denne deskriptive studien viser behov for videre forskningssamarbeid mellom studiestedene, for å utvikle en bioingeniørutdanning av best mulig kvalitet for fremtiden.publishedVersio

Praksisstudienes omfang og organisering i bioingeniørutdanningen

Since 2005, scope and content of the biomedical laboratory scientist education have been defined by national guidelines to ensure equal final competence for all candidates. From 2019, a new regulation on national guidelines was approved. Both new and old guidelines have regulated the learning outcomes and scope of practice in education. In this study, the scope and organization of the practical studies at the various study programs are characterized, in order to investigate differences and similarities and to facilitate that the study programs can learn from each other. Data has been obtained from each study program for the scope and organization of supervised internal practice, external practice and the bachelor thesis are reported for all study programs before and after the introduction of new guidelines. This study shows differences in the scope and organization of the practice and the bachelor thesis between the study programs in Norway and especially in the organization of internal practice. At the same time, the new guidelines open up for a further interpretation of the concept of practice and of requirements for the scope and organization of practice. In order to ensure equal final competence for the graduates, regardless of place of study, there is a need to agree to a greater extent on what provides sufficient scope and good quality of clinical practice in the study programs of biomedical laboratory scientists. This descriptive study shows the need for further research collaboration between the educational institutions, in order to develop a biomedical laboratory scientist education of the best possible quality for the future.publishedVersio

Praksisstudienes omfang og organisering i bioingeniørutdanningen

Siden 2005 har omfang og innhold i bioingeniørutdanningen vært definert av en nasjonal rammeplan for å sikre likeverdig sluttkompetanse for alle kandidater. Fra 2019 ble en ny forskrift om nasjonale retningslinjer fastsatt. Både tidligere rammeplan og nye retningslinjer har regulert læringsutbyttene og omfang av praksis i utdanningen. I denne studien kartlegges omfang og organisering av praksisstudiene ved de ulike studieprogrammene, dette for å undersøke forskjeller og likheter og tilrettelegge for at studiestedene kan lære av hverandre. Det er innhentet data fra hvert studieprogram for omfang og organisering av veiledet intern praksis, ekstern praksis samt bacheloroppgaven. Dette ble kartlagt for alle studiestedene før og etter innføring av nye retningslinjer. Resultatene viser ulikheter i omfang og organisering av praksisstudiene og bacheloroppgaven mellom studieprogrammene i Norge og da særlig i organisering av intern praksis. De nye retningslinjene åpner samtidig opp for en videre fortolkning av praksisbegrepet og av krav til omfang og organisering av praksis. For å sikre likeverdig sluttkompetanse for ferdigutdannede, uavhengig av studiested, er det behov for i større grad enes om hva som gir tilstrekkelig omfang og god kvalitet i praksis i bioingeniørutdanningen. Denne deskriptive studien viser behov for videre forskningssamarbeid mellom studiestedene, for å utvikle en bioingeniørutdanning av best mulig kvalitet for fremtiden.publishedVersio

Interaction engineered three-helix bundle domains for protein recovery and detection

HTML clipboard The great advances in DNA technology, e.g. sequencing and recombinant DNA techniques, have given us the genetic information and the tools needed to effectively produce recombinant proteins. Recombinant proteins are valuable means in biotechnological applications and are also emerging as alternatives in therapeutic applications. Traditionally, monoclonal antibodies have been the natural choice for biotechnological and therapeutic applications due to their ability to bind a huge range of different molecules and their natural good affinity. However, the large size of antibodies (150 kDa) limits tissue penetration and the recombinant expression is complicated. Therefore, alternative binders with smaller sizes have been derived from antibodies and alternative scaffolds. In this thesis, two structurally similar domains, Zbasic and ABDz1, have been used as purification tags in different contexts. They are both three-helical bundles and derived from bacterial surface domains, but share no sequence homology. Furthermore, by redesign of the scaffold used for ABDz1, a molecule intended for drug targeting with extended in-vivo half-life has been engineered. In Papers I and II, the poly-cationic tag Zbasic is explored and evaluated. Paper I describes the successful investigation of Zbasic as a purification handle under denaturating conditions. Moreover, Zbasic is evaluated as an interaction domain in matrixassisted refolding. Two different proteins were successfully refolded using the same setup without individual optimization. In Paper II, Zbasic is further explored as a purification handle under non-native conditions in a multi-parallel setup. In total, 22 proteins with varying characteristics are successfully purified using a multi-parallel protein purification protocol and a robotic system. Without modifications, the system can purify up to 60 proteins without manual handling. Paper I and II clearly demonstrate that Zbasic can be used as an interaction domain in matrix-assisted refolding and that it offers a good alternative to the commonly used His6-tag under denaturating conditions. In paper III, the small bifunctional ABDz1 is selected from a phage display library. Endowed with two different binding interfaces, ABDz1 is capable of binding both the HSA-sepharose and the protein A-derived MabSelect SuRe-matrix. The bifunctionality of the domain is exploited in an orthogonal affinity setup. Three target proteins are successfully purified using the HSA-matrix and the MabSelect SuRe-matrix. Furthermore, the purity of the target proteins is effectively improved by combining the two chromatographic steps. Thus, paper III shows that the small ABDz1 can be used as an effective purification handle and dual affinity tag without target specific optimization. Paper IV describes the selection and affinity maturation of small bispecific drug-targeting molecules. First generation binders against tumor necrosis factor-α are selected using phage display. Thereafter on-cell surface display and flow cytometry is used to select second-generation binders. The binding to tumor necrosis factor-α is improved up to 30 times as compared to the best first generation binder, and a 6-fold improvement of the binding strength was possible with retained HSA affinity. Paper III and IV clearly demonstrate that dual interaction surfaces can successfully be grafted on a small proteinaceous domain, and that the strategy in paper IV can be used for dual selection of bifunctional binders.QC20100610</p

To meet on young peoples terms in young peoples' social arena : - A qualitative study on youth workers experience of on line counselling of young people

With this research we aimed to seek how youth workers express their experience of counselling on the internet compared to traditional counselling face to face with a theoretical ground in semiotics and symbolic interactionism. Our questions were 1) How do youth workers describe their experiences from anonymous, faceless online counselling? 2) How do youth workers experience the differences between online counselling and the traditional face-to-face counselling? 3) Which possibilities and difficulties do they see in online communication with adolescents? We used semi-structed interviews with seven youth workers. Six of those are active in non-profit organisations, and one in social services. The results from our interviews showed that the interviewees saw that misunderstandings were more frequent on the internet, but that online counselling with adolescents still was easier than in the face-to-face meetings. To compensate for the non verbal part of communication in real life, symbols is seen as very important in online communication. The youth workers meant that the development of the on line counselling has opened a new arena. An arena where they get an opportunity to meet young people whom they don´t believe they would have met elsewhere. We found that this is a new, not fully developed culture with its own langue which we are about to learn. Once we understand and are capable of using this language new ways to meet with clients will be opened

High school students' sedentary and physical activity during school closure caused by Covid-19

Introduktion: Den 11 mars 2020 klassades viruset Covid-19 som en pandemi. För att bromsa smittspridningen beslutade regeringen att stänga gymnasieskolorna runt om i Sverige och övergå till fjärr- och distansundervisning. Flertal idrottsföreningar har inte heller kunnat bedriva sin verksamhet på grund av pandemin. Trots att restriktionerna har varit nödvändiga för att minska smittspridningen kan det ge konsekvenser på ungdomars och unga vuxnas rörelsevanor.  Syfte: Studera förekomst av stillasittande och fysisk aktivitet hos gymnasieungdomar i Västerbottens inland till följd av skolstängningen orsakat av Covid-19. Metod: Undersökningen utfördes bland 86 gymnasieelever (16–20 år) i Västerbottens inland. En webbenkät med 23 frågor skickades ut till tre gymnasieskolor i olika kommuner. Enkäten var tillgänglig mellan 2 februari och 16 februari 2021. Deskriptiv analys, Chi2-test och Man Whitney U användes för att analysera och jämföra data.  Resultat: I denna studie ansåg 81,4 % av eleverna att deras stillasittande hade ökat och 55,8 % ansåg att deras fysiska aktivitet minskat under pandemin. Totalt var det 55,8 % som inte uppnådde FYSS rekommendationer under Covid-19. Unga vuxna (18–20 år) och unga kvinnor visade en viss tendens att vara mindre fysiskt aktiva i jämförelse med ungdomar (16–17 år). Resultatet generade ingen statistisk signifikant skillnad avseende fysisk aktivitet och stillasittande mellan unga kvinnor och unga män. Valet av fysisk aktivitet förändrades under pandemin i jämförelse med innan.  Konklusion: Resultatet visar en negativ påverkan på stillasittande och fysisk aktivitet hos ungdomar och unga vuxna i Västerbottens inland på grund av pandemin. Detta skulle kunna leda till ökade hälsorisker hos ungdomar och unga vuxna i Västerbottens inland. Detta resultat kan vägleda insatser för att främja gymnasieelevers hälsa till följd av pandemin. Fortsatt forskning krävs då pandemin är pågående.

On the run the the land of border controls : A study of Dagens Nyheter and Sydsvenskan’s portrayal of refugees

In the autumn of 2015 a huge amount of refugees were on the run to Sweden. The news media described the situation as a refugee crisis and border controls were initiated. The aim of this study was to examine how two Swedish newspapers, one national and one local, portrayed refugees at the time when the border controls were initiated in Sweden. The questions examined were: How are the refugees portrayed, what are the similarities and differences between Dagens Nyheter and Sydsvenskan’s portrayal of the refugees? To approach this we made two different analysis of reports from each newspaper, three from Dagens Nyheter and two from Sydsvenskan. We did a framing analysis on the text and a semiotic analysis on the pictures. Findings show that the portrayal of refugees are very similar in both newspapers. The refugees are described as inferior, but at the same time brave, well-educated and hopeful. We also found that refugees often are portrayed as “the others” and the citizens of Sweden are portrayed as “we”. Although the similarities outweighs, we found some interesting differences in the newspapers portrayal of the refugees. For example, in Dagens Nyheter the hopefulness is combined with a positive and joyful approach, while Sydsvenskan describes the hope with an uncertainty and concern about the future

Introducing the Affinity Binder Knockdown Initiative—A public–private partnership for validation of affinity reagents

The newly launched Affinity Binder Knockdown Initiative encourages antibody suppliers and users to join this public–private partnership, which uses crowdsourcing to collect characterization data on antibodies. Researchers are asked to share validation data from experiments where gene-editing techniques (such as siRNA or CRISPR) have been used to verify antibody binding. The initiative is launched under the aegis of Antibodypedia, a database designed to allow comparisons and scoring of publicly available antibodies towards human protein targets. What is known about an antibody is the foundation of the scoring and ranking system in Antibodypedia