Bakgrund: Bioteknikbranschen är en bransch som präglas av stor osäkerhet. Många företags fortsatta existens beror på om företagets projekt eller produkt får godkänt av läkemedelsverket, vilket gör en värdering av bolaget under utvecklingsperioden mycket svår. Den vanligaste metoden vid värdering av bioteknikföretag idag är kassaflödesmetoden. Denna metod anses dock av vissa aktörer på den finansiella marknaden vara otillräcklig. Istället förespråkas en metod med hjälp av reala optioner som de anser tar bättre hänsyn till den osäkerhet som omgärdar bioteknikföretag. Syfte: Syftet med denna uppsats är att utveckla en real optionsmodell för värdering av Bioteknikföretag samt att pröva denna på ett fallföretag. Genomförande: För att uppfylla syftet utgicks från existerande företagsvärderingsmodeller och befintlig optionsteori. Vidare studerades reala optioner, och vilka av dessa som på ett bra sätt kunde förknippas med den osäkerhet som existerar i bioteknikföretag. Genom hela processen med utvecklandet av värderingsmodellen användes ett fallföretag, Diamyd Medical AB, och dess finansiella historia samt framtidsprognoser. Därutöver har information om bioteknikmarknaden och de förutsättningar företagen inom denna bransch ställs inför använts. Slutsats: Den skapade värderingsmodellen med hjälp av reala optioner visade sig på vårt fallföretag vara ett mycket användbart redskap och som på ett bra sätt klarar av att hantera den osäkerhet som omgärdar många bioteknikföretag och andra forskningsintensiva företag. / Background: The Biotech industry is an industry that is characterized by great uncertainty. Many companies future existence depends on if the company’s project or product gets an approval from the deciding authority, which makes a valuation of the company during the developing period very difficult. The most utilized method when valuing biotech firm today is the cash flow method. This method is nevertheless considered inadequate by some actors at the financial market. Instead a method using real options is recommended by some experts, which they think takes better concern to the uncertainty that surrounds biotech firms. Purpose: The purpose with this thesis is to develop a real option model to use when valuing biotech firm, and to try it on a case company. Implementation: To attain the goal for the thesis, the authors started out with the traditional methods of valuation and the existing theory surrounding financial options. Furthermore the authors studied real option and which of them that could be applied on the complex nature of biotechnology firms. Throughout the whole development process of the valuation model the authors used Diamyd Medical AB as a case company and in addition to that they also used more general information about the biotechnology sector. Conclusion: The valuation model containing real options proved to be a useful instrument to establish a monetary value on biotechnology firms.

In previous studies the carboxyl-terminus of microtubule-associated protein 1S was shown to interact with the N-methyl-D-aspartate receptor subunit NR3A (Eriksson et. al.)1.  In this study, interactions between three truncations of the microtubule-associated proteins 1B and one truncation of the microtubule-associated protein 1S carboxyl-terminus and the N-methyl-D-aspartate receptor subunits NR1 and NR3A were examined. The study showed that an interaction occurred between amino acids 2167 to 2365 of the microtubule-associated protein 1B and NR3A.  That region of microtubule associated protein 1B corresponds to a microtubule-binding region in the light chain. It has been shown in earlier studies (Reviewed in Halpain S. et a12, Riederer, BM.  et.al3.) that the light chain is a active part of the protein that have been post translational cleaved. The MAP 1 proteins are present in all tissue but has higher concentrations in the Post Synaptic Density of neurons in the central nervous system.  The N-methyl-D-aspartate receptors are present in glial cells and in the dendritic shafts of the central nervous system neurons (Eriksson et. al.)1 . The diseases were these proteins may play a part is mainly memory destructive diseases such as Alzheimers disease and in muscular dystrophy, but these assumptions are still being speculated.

The main objective of this investigation is to determine the effect of different physical and chemical parameters on the performance of biofilters, treating hydrophobic organic compounds. pinene was used as a model substance. Alpha-pinene is commonly found in wood, and therefore found at wood storage facilities and wood processing industries [1]. In this experiment two biofilter which were in equal size, were used. Both of them were filled with perlite for treating the alpha pinene contaminated air. One of the columns contained perlite partially coated with silicone oil to make the surface of perlite more hydrophobic. The filters were run at 5, 2.5 and 1.5 l min-1 air flow rate. The results showed that the silicone oil amended filter performed better at 2.5 l/min with a maximum removal rate of 20 g / (m3 h) in comparison with 15 g/ (m3 h) for the filter without oil. The efficiency was approximately the same for both filters at 1.5 and 5 l/min (40 compare to 35 g/m3 bed h). The flow rate was then set to 2.5 l/min once more. The results showed that the removal rate had increased to approximately 35 g/ (m3 h) and that the efficiency of both filters was approximately the same. The difference in results between the initial run and the later at 2.5 l/min is probably depending on that the microorganisms had become adapted to the α-pinene and that the microorganism communities developed differently in the two filters. / Syftet med denna studie var att undersöka om en ökning av biofilterytans hydrofobicitet kan förbättra effektiviteten med avseende på nedbrytning av en hydrofob förening såsom -pinen. En hydrofob yta skulle kunna medföra en ökning i adsorption av hydrofoba föroreningar vilket i sin tur skulle kunna öka tillgängligheten hos föroreningarna för mikroorganismerna. Försöken utfördes med hjälp av två biofilter fyllda med perlit. Ett filter fylldes med obehandlad perlit medan det andra filtret fylldes med perlit som blandats med silikonlja. Båda filtren ympades med mikroorganismer som växt på träflis. Dessa mikroorganismer är anpassade för att bryta ner -pinen då denna förening friges i stora mängder från trä. Resultaten visar att filtret innehållande perlit täckt med silikonolja var mer effektivt med avseende på nedbrytning avpinen under den första tiden av studien (de första 30 dagarna). Skillnaden i effektivitet mellan filtren minskade dock betydligt med tiden så att skillnaden inte var signifikant under resten av studien. Prover från vätskefasen från de båda filtren undersöktes i mikroskop. Dessa studier visade att olika mikroorganismer dominerade i de båda filtren. Filtret innehållande oljetäckt perlit dominerades av stavformiga bakterier medan filtret innehållande obehandlad perlit dominerades av kocker. Båda filtren innehöll en blandkultur av kocker och stavar. Dessutom växte svamp i båda filtren. Dessa resultat visar att en förändring av ytans hydrofobicitet inte bara kan leda till en ökad adsorption av förorening utan den kan också leda till att andra mikroorganismer får en dominerande roll i filtret. Den maximala nedbrytningshastigheten låg mellan 35 och 40 g pinen/(m3h) under hela studien förutom de 30 första dagarna oberoende av luftflödeshastighet (1,5 , 2,5 respektive 5 l/min). Den maximala nedbrytningshastigheten under inledningen av försöket var 15 g/(m3 h) för filtret innehållande obehandlad perlit och 20 g/(m3h) för filtret innehållande oljetäckt perlit. Luftflödet var 2,5 l/min under den inledande perioden. Ökningen i nedbrytningshastighet med tiden indikerar att mikroorganismerna anpassade sig till miljön. Den relativt konstanta nedbrytningshastigheten under resten av studien indikerar att det var de biokemiska reaktionerna som var hastighetsbegränsande och att masstransporten inte hade någon dominerande roll med avseende på att begränsa hastigheten.

Rapid wound healing is important to regain the skins protective function after injury. Studies have shown that enamel matrix proteins (EMP) have many desirable effects which may accelerate wound healing [Bosshardt et al. 2008]. Polymers (Polymer A, B and C) were formed into a mat form, with or without incorporated enamel matrix derivative (EMD) (Collaboration partner). The materials may be suitable for wound care and drug delivery systems. Protein release tests were performed on samples incubated in physiological-like solution using pyrogallol red staining, ultraviolet (UV) spectrophotometer and high-performance liquid chromatography (HPLC). Protein was detected in Polymer A material samples, compared to a reference material sample, using pyrogallol red staining. An in vitro experiment showed that normal human dermal fibroblasts (NHDF) cultivated with Polymer A material (with EMD) had significantly higher viability than NHDF cultivated with reference material (Polymer A without EMD) and comparable viability to fibroblasts grown with either 0.1 mg EMD in solution or with 10% fetal calf serum. Images taken of Polymer A material, with incorporated Fluorescein isothiocyanate- (FITC) labeled EMD, indicate a homogenous distribution of EMD peptides and/or EMD aggregates throughout the material. A dressing which contains an active substance may have clinical promise for wound care applications.

Stockholm Water is a water and sewage company with Henriksdal as one of two wastewater treatment plants (WWTPs). At Henriksdal wastewater sludge generated in the wastewater treatment process is digested which generate biogas; a mixture of mainly methane and carbon dioxide. If purified to methane content of 96 - 98 % this gas is called biomethane. Biogasmax is a project aiming to reduce the use of fossile fuels in Europe by providing that biogas is a good technical, economical and environmental alternative as vehicle fuel. The specific aim for Stockholm Water is to increase the biogas production at the existing plant in Henriksdal. Enzymatic treatment of wastewater sludge is an innovative technique earlier proofed to increase the biogas production from wastewater sludge with up to 60 %. The enzyme activity is in turn proven to significantly increase in the presence of a cation binding agent. One aim with this thesis was to investigate if the sludge from Henriksdal wastewater treatment process at all is affected of enzymatic treatment in presence of a cation binding agent since this has shown to have some significance. The chemical oxygen demand (COD) was measured in the liquid phase of sludge after treatment and used as a measurement of treatment effect. Another aim of this thesis was to look into the possibility to increase the methane production from sludge at Henriksdal WWTP. This was investigated through batch laboratory digestion tests. The sludge from Henriksdal WWTP was shown to be a good substrate for the enzymes added. COD in the liquid phase was increased with 17 – 32 % depending on the dose of enzymes and sodium citrate added. Digestion of sludge with a total addition of 18.6 mg enzymes per 1 g total solids (TS) and a concentration of 5 mM sodium citrate increased the methane production with almost 18 % compared to untreated sludge. This equals an increase of 18.3 % when converted to represent a totally blended and continuous digestion chamber at Henriksdal WWTP. The increased methane production also results in a sludge reduction out from the digestion chambers. The increased methane production and sludge reduction though does not fulfil the increased costs for the enzymes and sodium citrate applied. These doses must be decreased and the costs for both enzymes and sodium citrate must be reduced for this technique to be economically feasible in a full scale operation.

Many things might happen in the body when a titanium implant is inserted into bone. Examples are activation of the immune system and imbalance between bone formation and bone resorption, which might lead to damaged bone around the implant and at worse, loosening of the implant. Bisphosphonates, BP’s, is a class of drugs that is able to decrease the osteoclast (bone resorption cell) activity and thereby strengthen the bone. FibMat2.0 is a fibrinogen matrix and consists of a thin protein layer which can be applied on an implant and act as a local drug delivery system. The work in this thesis was divided into two parts where aim of the first part was to study FibMat2.0 with integrated BP’s, and their effect in the presence of blood. The aim for the second part was to determine whether it was possible to incorporate antithrombotic drugs into the fibrinogen matrix. No detection method for the amount of drugs incorporated into the fibrinogen matrix was used but the fact that the drugs gave effect was verifying that it is possible to integrate other drugs than BP’s into FibMat2.0. Methods that have been used in the experiments in presence of blood are imaging of coagulation, fluorescence microscopy and cone-and-plate. For the first part, the results showed that surfaces incubated with fibrinogen and fibrinogen with integrated BP’s act alike in regard to coagulation and platelet adhesion. Compared to titanium, which is known to be a biocompatible material, the surfaces with fibrinogen and fibrinogen with BP’s behave similar in regard to platelet adhesion. When it comes to coagulation, the surfaces coated with fibrinogen with or without an addition of BP’s have shown a longer coagulation time compared to the clean titanium surface. For the second part, some conclusions have been drawn according to the results. Heparin and hirudin have shown anticoagulant effects when integrated in the matrix. The platelet inhibitor cangrelor seemed to have better effect when added in blood and incubated compared to incubation with the platelet inhibitor on the surface before incubation in blood. Finally, when combining heparin and cangrelor, very clear differences in regard to formation of fibrin network could be seen. It seems promising to be able to load different kind of drugs in FibMat2.0.

Påväxt på båtar och andra föremål som sätts ner i vattnet är ett stort och kostsamt problem världen över. I Sverige orsakar havstulpanen, Balanus improvisius, den största påväxten. Havstulpanen har många förstadier innan den blir en vuxen havstulpan. I det stadiet innan den sätter sig fast på olika platser, kallas den cyprid. Det tar mellan tre dagar och upp till några veckor innan cypriden valt plats att settla på. Den väljer plats mycket noggrant då den skall tillbringa resterade delen av sitt liv på denna. Mycket forskning läggs ner på att finna ett alternativ till dagens giftiga båtbottenfärger. Tennorganiska föreningar, som använts för att motverka settling på båtbottnar, blir totalförbjudet 1 januari, 2008. Nu mer tillsätts allt oftare kopparföreningar istället vilket inte är någon större förbättring för miljön. Det finns idag många miljövänliga färger, som t.ex. silikonbaserad färg, men de fungerar inte lika bra som färgerna med metaller. Det forskas även på om ytan på båtbotten skall kunna förändras mekaniskt, för att motverka havstulpanens settling. De substanser som är testade i denna rapport är selenmetionin och nitroglycerin. De testades i olika koncentrationsintervaller och i olika blandningar med varandra. Med en förhoppning om att selenmetioninen binder in i föreningar och då att hastigheten för reaktioner med tioler kan minska drastiskt och leda till gifteffekter, och att nitroglycerinen skall pacificera cypriden genom att binda till hemoglobinet istället för syre, skall dessa två substanser hålla cypriderna borta från båtbottnarna. Med föraningar om att substanserna har en synergistisk effekt då de blandas, skall lägre koncentrationer kunna användas. Då för höga koncentrationer är kostsamt och har troligen större negativ påverkan på miljön. Selenmetionin visade en antydan till att minska settling men nitroglycerinen hade en mer främjande effekt på cypridens settling. Det visade sig även att substanserna hade en synergistisk effekt och därmed kan koncentrationerna för substanserna sänkas. / Biological growth on boats and other objects that are placed in water is a big and expensive problem over the entire world. In Sweden, the barnacle, Balanus improvisius, grows the most easily. This barnacle has many larvae stages before it becomes mature. During the larvae stage when it searching for a surface to settle on it is called cyprid. It takes between three days and a couple of weeks before a cyprid chooses a spot to settle on. It chooses the spot to settle on very carefully because it will spend the rest of its life on that exact place. A lot of research goes into to finding a good alternative to today’s toxic boat paints. Tin organic compounds, as used to counteract settling of cyprids on boats, will be totally forbidden in January, 2008. As a replacement, copper compounds are being added more often to paints. These are not much better for the environment. Today there are many environmentally friendly paints, as for example silicon-based paint, but they do not work as well as the paints with metals. Research is also looking at mechanically changing a boat’s surface so that barnacles cannot attach as easily. The substances that are tested in this report are selenomethionine and nitro glycerine. They were tested in different concentration intervals and in different mixes with each other with the hope of that selenomethionine will bound into compounds and then the reaction velocity with thioles will be reduced and give toxic effects. The nitro glycerine will make the barnacle passive through the idea that the NO is binding into a heme group instead of oxygen. It was hoped that the substances show a synergistic effect when they are mixed, allowing lower concentrations to be used. With too high concentrations in the paint, the paint becomes too expensive and likely has a bigger negative influence on the environment. Selenomethionine showed an indication to reduce the likelihood that barnacles would settle on a pained surface, but nitro glycerine, on the other hand, caused an increased rate of cyprid settling. It was also shown that the substances had a synergistic effect and thus the concentrations for the substances can be lowered.

Silver was used for its antimicrobial effect by the ancient Greeks, long before the existence of microorganisms were first suspected. Nowadays a wide range of antimicrobial dressings containing silver, either incorporated within or applied on the dressings, are available for clinical use. This type of dressings is designed to provide the antimicrobial activity of silver in a more convenient application. The aim with this master thesis was to evaluate if silver release and antimicrobial effect of nine silver containing dressings are dependent on the test medium and if there is any relation between silver release and antimicrobial effect. Release of silver and antimicrobial effect was evaluated by using a 6-well co-culture system, with inoculated test medium in the wells and dressing pieces in the culture well inserts. Three different test media with increased complexity and nutrient value were inoculated with either Results show that release of silver depends on the test fluid used; for phosphate buffered saline (PBS), the silver concentration was as most 1.2 ppm, but for a complex media containing calf serum (SWF), it varied from 9 ppm to 134 ppm. The viable counts in PBS were reduced by at least 3 log units for all dressings and bacteria, whereas in SWF there were no reduction and instead growth was observed. In general, a high release resulted in less bacterial growth. Results also indicated that kinetics of silver release affect the antimicrobial effect. It is likely to assume that it is important for a dressing to release silver quickly. It has previously not been possible to correlate silver release of wound care dressings and antimicrobial effect, since the two factors have been measured in different test systems and in different media. Since both factors depend on test medium and method used, it is shown in the present study that it is important to use relevant test medium for in-vitro evaluation. When measuring silver release and antimicrobial effect in the same test system, a relation is found.

<p>The important protein hormone insulin, responsible for different kind of functions in our body but mainly storage of nutrients, has for a long time been used for treatment of diabetic patients. This important protein is both physically and chemically unstable. Especially during production where the insulin protein is exposed to unnatural environmental conditions such as acidic pH has this been causing problems since huge volumes of the product go to waste.</p><p>In the human body the environment for the protein is tolerable with normal body temperature and the right pH, but when the protein is commercially synthesised the environmental conditions are not ultimate. What happens during these unfavourable conditions is that the insulin starts to fibrillate. Meaning that linear, biologically inactive aggregates are formed. If then under these kinds of conditions such as high temperature and acidic pH, the insulin comes in contact with hydrophobic surfaces then the fibrillation of the protein goes even faster.</p><p>In the following experiment I am going to investigate if the experiments and conclusions done before, where different kinds of additives to insulin solutions have been used to enhance the amyloid fibrillation of insulin, are as effective as it has been proposed and I am going to prove that the presence of hydrophobic surfaces, such as coated silicon surfaces or glass and addition of preformed fibrils, so called seeds, increase amyloid fibrillation of the insulin protein under certain conditions, in comparison with the normal fibrillation under the same conditions.</p>

<p>A capture step with protein A is the most common purification step in the downstream purification process of monoclonal antibodies. It is therefore of great importance to increase the knowledge of the interactions involved in this purification technique. The purpose of this master thesis project was to determine the affinity of protein A domains to IgG subclasses by surface plasmon resonance (SPR).</p><p>Besides the five homologous IgG-binding protein A domains (E, D, A, B, and C) an engineered domain, similar to domain B and used in the protein A media MabSelect Sure™ (GE Healthcare) was included in the study. The domains were expressed in E.coli, affinity purified and immobilized onto sensor chip surfaces by amine coupling. The antibodies used in the interaction analyses were of the human IgG subclasses 1, 2, 3, and 4. Affinity determination was performed by kinetic analyses with the SPR-biosensor Biacore™ 2000.</p><p>All human IgG subclasses except IgG3 were shown to bind to all protein A domains including the monomer of the SuRe ligand. The equilibrium constants, KD-values, obtained were all in the low nanomolar range. For IgG1 and IgG4, no significantly differences in the affinity to any of the protein A domains were found, except for domain E where there might be quality issues of the prepared domain. Furthermore, a detected quality issue with the commercial IgG2 made it impossible to determine the KD-values for this subclass with any reliability.</p>