Methods of genetic diversity creation and functional display for directed evolution experiments

Protein engineering aims to improve the properties of enzymes and affinity reagents by genetic changes. Typical engineered properties are affinity, specificity, stability, expression, and solubility. Because proteins are complex biomolecules, the effects of specific genetic changes are seldom predictable. Consequently, a popular strategy in protein engineering is to create a library of genetic variants of the target molecule, and render the population in a selection process to sort the variants by the desired property. This technique, called directed evolution, is a central tool for trimming protein-based products used in a wide range of applications from laundry detergents to anti-cancer drugs. New methods are continuously needed to generate larger gene repertoires and compatible selection platforms to shorten the development timeline for new biochemicals. In the first study of this thesis, primer extension mutagenesis was revisited to establish higher quality gene variant libraries in Escherichia coli cells. In the second study, recombination was explored as a method to expand the number of screenable enzyme variants. A selection platform was developed to improve antigen binding fragment (Fab) display on filamentous phages in the third article and, in the fourth study, novel design concepts were tested by two differentially randomized recombinant antibody libraries. Finally, in the last study, the performance of the same antibody repertoire was compared in phage display selections as a genetic fusion to different phage capsid proteins and in different antibody formats, Fab vs. single chain variable fragment (ScFv), in order to find out the most suitable display platform for the library at hand. As a result of the studies, a novel gene library construction method, termed selective rolling circle amplification (sRCA), was developed. The method increases mutagenesis frequency close to 100% in the final library and the number of transformants over 100-fold compared to traditional primer extension mutagenesis. In the second study, Cre/loxP recombination was found to be an appropriate tool to resolve the DNA concatemer resulting from error-prone RCA (epRCA) mutagenesis into monomeric circular DNA units for higher efficiency transformation into E. coli. Library selections against antigens of various size in the fourth study demonstrated that diversity placed closer to the antigen binding site of antibodies supports generation of antibodies against haptens and peptides, whereas diversity at more peripheral locations is better suited for targeting proteins. The conclusion from a comparison of the display formats was that truncated capsid protein three (p3Δ) of filamentous phage was superior to the full-length p3 and protein nine (p9) in obtaining a high number of uniquely specific clones. Especially for digoxigenin, a difficult hapten target, the antibody repertoire as ScFv-p3Δ provided the clones with the highest affinity for binding. This thesis on the construction, design, and selection of gene variant libraries contributes to the practical know-how in directed evolution and contains useful information for scientists in the field to support their undertakings.

Simulation of new p-type pixel strip detector with enhanced multiplication effect

This Master’s Thesis is dedicated to the simulation of new p-type pixel strip detector with enhanced multiplication effect. It is done for high-energy physics experiments upgrade such as Super Large Hadron Collider especially for Compact Muon Solenoid particle track silicon detectors. These detectors are used in very harsh radiation environment and should have good radiation hardness. The device engineering technology for developing more radiation hard particle detectors is used for minimizing the radiation degradation. New detector structure with enhanced multiplication effect is proposed in this work. There are studies of electric field and electric charge distribution of conventional and new p-type detector under reverse voltage bias and irradiation. Finally, the dependence of the anode current from the applied cathode reverse voltage bias under irradiation is obtained in this Thesis. For simulation Silvaco Technology Computer Aided Design software was used. Athena was used for creation of doping profiles and device structures and Atlas was used for getting electrical characteristics of the studied devices. The program codes for this software are represented in Appendixes.

Particle model for simulating limestone reactions in novel fluidised bed energy applications

The development of carbon capture and storage (CCS) has raised interest towards novel fluidised bed (FB) energy applications. In these applications, limestone can be utilized for S02 and/or CO2 capture. The conditions in the new applications differ from the traditional atmospheric and pressurised circulating fluidised bed (CFB) combustion conditions in which the limestone is successfully used for SO2 capture. In this work, a detailed physical single particle model with a description of the mass and energy transfer inside the particle for limestone was developed. The novelty of this model was to take into account the simultaneous reactions, changing conditions, and the effect of advection. Especially, the capability to study the cyclic behaviour of limestone on both sides of the calcination-carbonation equilibrium curve is important in the novel conditions. The significances of including advection or assuming diffusion control were studied in calcination. Especially, the effect of advection in calcination reaction in the novel combustion atmosphere was shown. The model was tested against experimental data; sulphur capture was studied in a laboratory reactor in different fluidised bed conditions. Different Conversion levels and sulphation patterns were examined in different atmospheres for one limestone type. The Conversion curves were well predicted with the model, and the mechanisms leading to the Conversion patterns were explained with the model simulations. In this work, it was also evaluated whether the transient environment has an effect on the limestone behaviour compared to the averaged conditions and in which conditions the effect is the largest. The difference between the averaged and transient conditions was notable only in the conditions which were close to the calcination-carbonation equilibrium curve. The results of this study suggest that the development of a simplified particle model requires a proper understanding of physical and chemical processes taking place in the particle during the reactions. The results of the study will be required when analysing complex limestone reaction phenomena or when developing the description of limestone behaviour in comprehensive 3D process models. In order to transfer the experimental observations to furnace conditions, the relevant mechanisms that take place need to be understood before the important ones can be selected for 3D process model. This study revealed the sulphur capture behaviour under transient oxy-fuel conditions, which is important when the oxy-fuel CFB process and process model are developed.

Value co-creation in gaming the industry: A qualitative research on virtual discussion forums

This study discusses the procedures of value co-creation that persist in gaming industry. The purpose of this study was to identify the procedures that persist in current video gaming industry which answers the main research problem how value is co-created in video gaming industry followed by three sub questions: (i) What is value co-creation in gaming industry? (ii) Who participates in value co-creation in gaming industry? (iii) What are the procedures that are involved in value co-creation in gaming industry? The theoretical background of the study consists of literature relating to the theory of marketing i.e., notion of value, conventional understanding of value creation, value chain, co-creation approach, co-production approach. The research adopted qualitative research approach. As a platform of relationship researcher used web 2.0 tool interface. Data were collected from the social networks and netnography method was applied for analyzing them. Findings show that customer and company both co-create optimum level of value while they interact with each other and within the customers as well. However mostly the C2C interaction, discussions and dialogues threads that emerged around the main discussion facilitated to co-create value. In this manner, companies require exploiting and further motivating, developing and supporting the interactions between customers participating in value creation. Hierarchy of value co-creation processes is the result derived from the identified challenges of value co-creation approach and discussion forums data analysis. Overall three general sets and seven topics were found that explored the phenomenon of customer to customer (C2C) and business to customer (B2C) interaction/debating for value co-creation through user generated contents. These topics describe how gamer contributes and interacts in co-creating value along with companies. A methodical quest in current research literature acknowledged numerous evolving flows of value in this study. These are general management perspective, new product development and innovation, virtual customer environment, service science and service dominant logic. Overall the topics deliver various realistic and conceptual implications for using and handling gamers in social networks for augmenting customers’ value co-creation process.

Supporting the creation, management, and long-term preservation of social science research data

Poster at Open Repositories 2014, Helsinki, Finland, June 9-13, 2014

Let's do data research work: the creation of a portal with research information from Catalan Universities

Presentation at Open Repositories 2014, Helsinki, Finland, June 9-13, 2014

Railways, Airports and Sea Container Operators as Publicly Listed Companies – Financial Performance and Shareholder Value Creation Perspective

Logistics infrastructure and transportation services have been the liability of countries and governments for decades, or these have been under strict regulation policies. One of the first branches opened for competition in EU as well as in other continents, has been air transports (operators, like passenger and freight) and road transports. These have resulted on lower costs, better connectivity and in most of the cases higher service quality. However, quite large amount of other logistics related activities are still directly (or indirectly) under governmental influence, e.g. railway infrastructure, road infrastructure, railway operations, airports, and sea ports. Due to the globalization, governmental influence is not that necessary in this sector, since transportation needs have increased with much more significant phase as compared to economic growth. Also freight transportation needs do not correlate with passenger side, due to the reason that only small number of areas in the world have specialized in the production of particular goods. Therefore, in number of cases public-private partnership, or even privately owned companies operating in these sub-branches have been identified as beneficial for countries, customers and further economic growth. The objective of this research work is to shed more light on these kinds of experiments, especially in the relatively unknown sub-branches of logistics like railways, airports and sea container transports. In this research work we have selected companies having public listed status in some stock exchange, and have needed amount of financial scale to be considered as serious company rather than start-up phase venture. Our research results show that railways and airports usually need high fixed investments, but have showed in the last five years generally good financial performance, both in terms of profitability and cash flow. In contrary to common belief of prosperity in globally growing container transports, sea vessel operators of containers have not shown that impressive financial performance. Generally margins in this business are thin, and profitability has been sacrificed in front of high growth – this also concerns cash flow performance, which has been lower too. However, as we examine these three logistics sub-branches through shareholder value development angle during time period of 2002-2007, we were surprised to find out that all of these three have outperformed general stock market indexes in this period. More surprising is the result that financially a bit less performing sea container transportation sector shows highest shareholder value gain in the examination period. Thus, it should be remembered that provided analysis shows only limited picture, since e.g. dividends were not taken into consideration in this research work. Therefore, e.g. US railway operators have disadvantage to other in the analysis, since they have been able to provide dividends for shareholders in long period of time. Based on this research work we argue that investment on transportation/logistics sector seems to be safe alternative, which yields with relatively low risk high gain. Although global economy would face smaller growth period, this sector seems to provide opportunities in more demanding situation as well.

Container logistic innovations in forest-energy sector: Markets, future service concepts and technical improvements

The report presents the results of the commercialization project called the Container logistic services for forest bioenergy. The project promotes new business that is emerging around overall container logistic services in the bioenergy sector. The results assess the European markets of the container logistics for biomass, enablers for new business creation and required service bundles for the concept. We also demonstrate the customer value of the container logistic services for different market segments. The concept analysis is based on concept mapping, quality function deployment process (QFD) and business network analysis. The business network analysis assesses key shareholders and their mutual connections. The performance of the roadside chipping chain is analysed by the logistic cost simulation, RFID system demonstration and freezing tests. The EU has set the renewable energy target to 20 % in 2020 of which Biomass could account for two-thirds. In the Europe, the production of wood fuels was 132.9 million solid-m3 in 2012 and production of wood chips and particles was 69.0 million solidm3. The wood-based chips and particle flows are suitable for container transportation providing market of 180.6 million loose- m3 which mean 4.5 million container loads per year. The intermodal logistics of trucks and trains are promising for the composite containers because the biomass does not freeze onto the inner surfaces in the unloading situations. The overall service concept includes several packages: container rental, container maintenance, terminal services, RFID-tracking service, and simulation and ERP-integration service. The container rental and maintenance would provide transportation entrepreneurs a way to increase the capacity without high investment costs. The RFID-concept would lead to better work planning improving profitability throughout the logistic chain and simulation supports fuel supply optimization.

User-generated experiences: how automated platforms for content creation and sharing has transformed information access

Esitys KDK-käytettävyystyöryhmän järjestämässä seminaarissa: Miten käyttäjien toiveet haastavat metatietokäytäntöjämme? / How users' expectations challenge our metadata practices? 30.9.2014.

Value Creation in Crowd-Business: Developing a Framework for Successful Implementation of Crowdsourcing Initiatives

By recent years the phenomenon called crowdsourcing has been acknowledged as an innovative form of value creation that must be taken seriously. Crowdsourcing can be defined as an act of outsourcing tasks originally performed inside an organization, or assigned externally in form of a business relationship, to an undefinably large, heterogeneous mass of potential actors. This thesis constructs a framework for successful implementation of crowdsourcing initiatives. Firms that rely entirely on their own research and ideas cannot compete with the innovative capacity that crowd-powered firms have. Nowadays, crowdsourcing has become one of the key capabilities of businesses due to its innovative capabilities, in addition to the existing internal resources of the firm. By utilizing crowdsourcing the business gains access to an enormous pool of competence and knowledge. However, various risks remain such as uncertainty of crowd structure and loss of internal know-how. Crowdsourcing Success Framework introduces a step by step model for implementing crowdsourcing into the everyday operations of the business. It starts from the decision to utilize crowdsourcing and continues further into planning, organizing and execution. Finally, this thesis presents the success factors of crowdsourcing initiative.