Antibody Development - Examples from viral and proteomic studies

This thesis, which is based on four original papers, covers different aspects of antibody development. The overall goal of my work has been to obtain an increased understanding of antibody repertoires as they develop in vivo and in vitro. In Paper I-III antibody repertoires was evaluated and used to dissect virus-induced antibody responses and characterize such antibodies. Furthermore, tools for more efficient studies of such characterized antibodies were developed. The study of antibody repertoires in this thesis reveals, not only insights of the composition of antibodies induced by a common virus, but also features for such binders developed by combinatorial and non-combinatorial approaches. Such developed antibodies may contribute with data important for future design of vaccine candidates. In Paper IV, limitations in biomarker verification processes are addressed. A proof-of-concept study was developed, describing an approach with high-throughput potential relying on enrichment of proteins by antibody fragments in complex serum samples followed by an MRM mass spectrometry readout. Taken together, the in vitro developed binders in combination with MRM readout provides a powerful method to quantify protein biomarkers in a high-throughput manner approaching the criteria for clinical usage

Förbättring av varmvattenproduktion i inomhusmodulen EcoZenith i360

Det blir allt viktigare med effektiv resurs- och energianvändning. Vatten är en av de viktigasteresurserna som finns och FN har till och med satt upp globala mål för att försöka öka tillgångentill rent vatten för hela jordens befolkning. I Sverige är det inte ovanligt att vatten slösas utifrån prioritering för komfort och bekvämlighet. Ett exempel är att låta duschen rinna för att få en varm och jämn temperatur innan duschningen påbörjas. Detta examensarbete görs i samarbetemed företaget CTC och berör varmvattenproduktion i deras produkt EcoZenith i360. Processen som regleras är värmeväxlingen mellan två vätskor i produktens plattvärmeväxlare. Syftet är att förbättra regleringen av varmvattentemperaturen med mål att utgående temperatur stabiliseras snabbt vid start. Målen är att minska regleringens insvängningstid, översväng och undersväng genom att införa information om aktuellt flöde för att anpassa regleringen samt anpassa det fasta startvärdet som används av cirkulationspumpen. För att kunna genomföra projektet behöver ett system skapas som hanterar kommunikationen med modulen och styrpumphastigheten. Ett krav på kommunikationen var att Modbusprotokoll skulle användas. Syftet har försökt uppnås genom att kartlägga den befintliga produkten för att hitta vilka faktorer som främst påverkar regleringen. Dynamikvariationer visades vara en avgörande faktor som föranledde att parameterstyrning med anpassade PID-regulator inställningar valdes. Kodprogram för den nya regleringen togs fram som hanterade dataöverföring med två olika slavar över en och samma kanal. Fel i dataöverföringen behövdes kunna detekteras och hanteras för att inte störa eller stanna upp regleringen. Resultatet av arbetet är att undersvängen i samtliga fall minskats medan insvängningstid och översväng har blandade resultat. Att minska undersvängen blev en prioriterad fråga i arbetet eftersom det till största del påverkar kundupplevelsen. Vid ett beräknat exempel uppskattas en minskning på 14 % av en användares dagliga vattenförbrukning kunnat uppnås genom användandet av den nya, framtagna regleringen.The importance of managing one’s use of natural resources and energy consumption is increasing. Water is one of the most important resources there is and the UN has set global goals to try and increase access to clean water for all mankind. In Sweden, it is not unheard of water being wasted due to a priority of comfort and convenience. One example is to let the water in the shower run before entering to make sure the temperature is hot and even. This thesis is made together with the company CTC and handles hot water production in their product EcoZenith i360. The process to be controlled is the heat exchange between two fluids inthe product's plate heat exchanger. The purpose is to improve the control of the hot water temperature in regards to making the produced water temperature quickly stabilize from the start. The goals are to decrease the settling time, overshoot, and undershoot for the process by adding information about actual water flow to the controller and adjusting the fixed pump speed. A system needs to be created to handle the communication between the module and other devices as well as being able to control the pump speed. One limitation is to use the Modbus protocol when implementing the communication. To achieve the thesis’ purpose the current module has been mapped out to identify what factors most affect the controller. Due to the dynamic range being the most significant factor, gain scheduling with fitted PID controller settings was chosen. A program with the new control was made to handle communication with two slaves over the same line. Error detection needed to be implemented and handled to ensure the system would not stop running. The result of the project showed that the undershoot decreased in all cases while the overshoot and settling time had various results. The undershoot became a prioritized goal since it mostly affected the customer experience. A calculated example shows a potential decrease in a user's daily water usage by 14 % if the new regulation is used

The relationship between ICA AB and Royal Ahold -A study on international ownership

Problem background: The 1990’s was a period of internationalisation within the retail sector. It was almost necessary to have a growth strategy with an international focus to be competitive on the retail market. The Swedish market-leading retail company ICA AB adapted to the increased internationalisation and developed an internationalisation strategy. In year 2000 the large Dutch international retail group Royal Ahold acquired 50 per cent of the shares in ICA AB. In connection with the transaction a shareholder agreement was established that was valid for 40 years and regulated the influence and control in ICA AB. In March of year 2013, Ahold divested its holdings in ICA AB to Hakon Invest, which is the financial organ within the ICA organisation. This lead to that ICA AB today is owned solely by Swedish shareholders. Problem discussion and objective: The objective of this thesis is to gain deeper knowledge in how two large companies within the retail industry corps with questions related to control and international partnership. This particular business deal evoked interest because of the interesting ownership structure, both within ICA AB and the relationship between ICA and Ahold. Deeper research into why Ahold became a part owner of ICA AB in year 2000, and why Ahold decided to sell the ICA ABshares in year 2013 is the foundation for the problem discussion in this thesis. Further investigations are made on how a business relationship like this originate and how and why is it terminated, and why a typical Swedish company decides to join partnership with a large foreign player. Method: Personal interviews have been conducted for the understanding of the problem discussion from the respondents’ perspective. Without the personal interviews the authors believe that it would not have been possible to reach the level of insight that was necessary for the quality of this thesis. Therefore a qualitative method was applied and part of the theoretical framework was developed after the interviews, which indicates that an inductive method was used to approach the objective. Observations: The observations are based on the four conducted interviews. The observations are divided according to a chronological order and important events to provide a pedagogic understanding of the content. Conclusion: Both companies had understandable motives to start the cooperation in year 2000 mainly due to the business environment at that time. The cooperation involved a change within the ICA organisation, mainly within the management but it was a notable fact for all parts of the organisation. The decision to end the relationship in year 2013 was mutual and a result of the new business strategy of Ahold in combination with ICA HF’s mission to secure the ICA-idea. Our conclusion is that this will benefit the ICA-idea and that the business areas that were not developed to its full potential during the time with Ahold now have the possibility to grow. An interesting question is how the listing of ICA Gruppen will affect the strategy of the company. Suggestions for further research: This thesis was written during the period of time directly after the buy-back of the shares of ICA AB. Therefore it would be interesting to investigate how the new ownership structure is going to affect the control over ICA AB and whether the recent transaction was beneficial or not for ICA AB. Since this thesis has a synoptically perspective it is possible to conduct deeper research into the ownership structure within ICA. Schuster (1989) examined the ownership relations within ICA from an agency theory in year 1989 but many structural changes have occurred since that period of time. Therefore it would be motivated to conduct a similar study, in particular after the most recent structural change

If I could download clothes, I would

Sammanfattning Titel: If I could download clothes, I would - En studie kring vad som påverkar konsumenters köpbeslut mellan e-handel och butikshandel. Författare: Fredrika Carlsson och Malin Cederberg Kurs: KSKK01 Service Management: Examensarbete för kandidatexamen Institution: Institutionen Service Management - Hotell och Restaurant, Campus Helsingborg, Lunds Universitet Handledare: Mia Larson Problembeskrivning: I samband med den växande e-handeln väljer fler konsumenter att överge butikshandeln och istället vända sig till e-handel. E-handeln och butikshandeln har både för och nackdelar, frågan är hur konsumenterna gör sitt val mellan de båda. Fokus i denna studie ligger på kläd- och filmuthyrningsbranschen och detta för att se om det går att härleda skillnader även mellan olika produkter och branscher. Syfte: Vårt syfte med denna studie är att undersöka vad som påverkar konsumenter i valet mellan e-handel och butikshandel. Metod: Studien baseras både på en kvantitativ samt en kvalitativ metodansats med utgångspunkt i enkätundersökning samt djupintervjuer. Slutsats: Vilken typ av produkt konsumenten vill införskaffa påverkar valet av tjänsteleverantör. De risker som är kopplade till e-handel och butikshandel påverkas även de av den produkt som ska konsumeras. Konsumenten väljer mellan de båda tjänsteleverantörerna utifrån ett individ-, produkt samt situationsbaserat förhållningssätt

Rällsögårdens missbruksprogram : En studie om klienter och personals erfarenheter av programmet och dess genomförande

The aim is that on the basis of interviews with personnel and clients examine which experiences they have of the drug abuse programme and its implementation. To be able to answer the aim following question has been formulated: (1) What do the personnel and client think about the drug abuse program arrangement? (2) In which extend is the program implemented as thought? (3) How do the personnel assesses the client’s participation? (4) How does the client’s think about their own participation in the program? (5) Do the client’s think that the program have been helpful in their personal development? A qualitative method was chosen and has used a semi structured interview. Total has ten interviews been implemented on Rällsögården, five with clients and five with personnel. The study's interpretation framework builds on research around drug abuse treatment and theories and methods that lie to shallow for the drug abuse programme. In the study part's assessments of the programme has as a whole been done, treatment integrity has been examined on the basis of factors such as education and implementation. Finally, have in some mean examined motivation within the clients on the basis of theirs and the personnel's views. One of the results that arrived in the study, where that an all in all assessment from the clients around the drug abuse programme where very difficult to get hold off in certain respects. This depended to a large extent on that the clients bar with itself experiences from earlier treatments with another treatment philosophy. The result has influenced the study as a whole. The study's head aim where that on the basis of Rällsögårdens commissions, try to understand how the drug abuse programme is experienced of the clients and the personnel on the basis of their different perspectives. The results show that the personnel in most cases has the education, motivation and commitments that is needed in order to do something good with the programme. Also the manual that is used has support in research where it is to be considered to give possibility to effective treatment. On the basis of this the conclusion can be drawn that the basic requirements are there, to carry out a good treatment programme. The results show however on several conditions that can influence the intentions negative

A dimerized single-chain variable fragment system for the assessment of neutralizing activity of phage display-selected antibody fragments specific for cytomegalovirus.

Abstract in UndeterminedCytomegalovirus (CMV) causes severe sequelae in congenitally infected newborns and may cause life-threatening disease in immuno-deficient patients. Recent findings demonstrate the possibility to alleviate the disease by infusing intravenous immunoglobulin G (IgG) preparations, indicating that antibodies are an effective therapeutic option. Modern molecular methodologies, like phage display, allow for the development of specific antibodies targeting virtually any antigen, including those of CMV. However, such methodologies do not in general result in products that by themselves mediate biological activity. To facilitate a semi-high-throughput approach for functional screening in future efforts to develop efficacious antibodies against CMV, we have integrated two different approaches to circumvent potential bottlenecks in such efforts. Firstly, we explored an approach that permits easy transfer of antibody fragment encoding genes from commonly used phage display vectors into vectors for the production of divalent immunoglobulins. Secondly, we demonstrate that such proteins can be applied in a novel reporter-based neutralization assay to establish a proof-of-concept workflow for the generation of neutralizing antibodies against CMV. We validated our approach by showing that divalent antibodies raised against the antigenic domain (AD)-2 region of gB effectively neutralized three different CMV strains (AD169, Towne and TB40/E), whereas two antibodies against the AD-1 region of gB displayed minor neutralizing capabilities. In conclusion, the methods investigated in this proof-of-concept study enables for a semi-high-throughput workflow in the screening and investigation of biological active antibodies

Humoral immunity targeting site I of antigenic domain 2 of glycoprotein B upon immunization with different cytomegalovirus candidate vaccines.

Glycoprotein B (gB) is a major component in several vaccines that are under development for prevention of disease by cytomegalovirus. It contains multiple determinants that are targets for neutralizing antibodies. One of them is site I of antigenic domain 2 (AD-2). The epitope, defined by short peptides, is quite conserved between different isolates. However, it is poorly immunogenic in natural infection. In this study we investigated the extent to which different vaccines, attenuated live Towne vaccine with or without priming with a canarypox virus coding for gB, or a recombinant gB vaccine adjuvanted with MF59, induced antibodies to this epitope. As in natural infection only a fraction of all subjects developed antibody responses against site I of AD-2 following vaccination. We suggest that strategies that enhance immunogenicity of this epitope will improve vaccine efficacy