Generation of affinity maturation libraries of PSMA targeting affibody molecules and selections to find improved binders

Prostate specific membrane antigen (PSMA) is a membrane protein expressed in both prostate cancer cells and in the neovasculature of many other solid tumors. It can act as a therapeutic target and as a target for an imaging agent for various cancers. Earlier, three different PSMA targeting affinity molecules, called Affibody molecules, were generated through phage selection. The aim of this thesis was to produce three corresponding affinity maturation libraries based on the three potential PSMA binders and select for improved binders through phage display. The affinity maturation libraries were produced, and two to four selection cycles were performed with yet insufficient enrichment. Therefore, more cycles must be performed in order to discover potential PSMA-binders with sufficient affinity.Prostataspecifikt membranantigen (PSMA) är ett membranprotein som uttrycks i både prostatacancerceller såväl som i nybildade blodkärl kring en tumör. Cancercellers uttryck av PSMA kan användas för att både behandla och upptäcka tumörer i kroppen. Tidigare har tre PSMA-sökande affinitetsmolekyler tagits fram, kallade Affibodymolekyler, genom fagdisplay. Syftet med detta examensarbete var att producera tre korresponderande affinitetsmatureringsbibliotek baserat på de tidigare potentiella PSMA-bindarna, och att selektera via fagdisplay för att hitta bindare med högre affinitet. De tre affinitetsmatureringsbiblioteken producerades och två till fyra selektionscykler har genomförts utan att se tillräcklig anrikning av bindare. Därför måste fler cykler genomföras i syfte att hitta potentiella PSMA-bindare med tillräckligt god affinitet

Generation of affinity maturation libraries of PSMA targeting affibody molecules and selections to find improved binders

Prostate specific membrane antigen (PSMA) is a membrane protein expressed in both prostate cancer cells and in the neovasculature of many other solid tumors. It can act as a therapeutic target and as a target for an imaging agent for various cancers. Earlier, three different PSMA targeting affinity molecules, called Affibody molecules, were generated through phage selection. The aim of this thesis was to produce three corresponding affinity maturation libraries based on the three potential PSMA binders and select for improved binders through phage display. The affinity maturation libraries were produced, and two to four selection cycles were performed with yet insufficient enrichment. Therefore, more cycles must be performed in order to discover potential PSMA-binders with sufficient affinity.Prostataspecifikt membranantigen (PSMA) är ett membranprotein som uttrycks i både prostatacancerceller såväl som i nybildade blodkärl kring en tumör. Cancercellers uttryck av PSMA kan användas för att både behandla och upptäcka tumörer i kroppen. Tidigare har tre PSMA-sökande affinitetsmolekyler tagits fram, kallade Affibodymolekyler, genom fagdisplay. Syftet med detta examensarbete var att producera tre korresponderande affinitetsmatureringsbibliotek baserat på de tidigare potentiella PSMA-bindarna, och att selektera via fagdisplay för att hitta bindare med högre affinitet. De tre affinitetsmatureringsbiblioteken producerades och två till fyra selektionscykler har genomförts utan att se tillräcklig anrikning av bindare. Därför måste fler cykler genomföras i syfte att hitta potentiella PSMA-bindare med tillräckligt god affinitet

Generation of affinity maturation libraries of PSMA targeting affibody molecules and selections to find improved binders

Prostate specific membrane antigen (PSMA) is a membrane protein expressed in both prostate cancer cells and in the neovasculature of many other solid tumors. It can act as a therapeutic target and as a target for an imaging agent for various cancers. Earlier, three different PSMA targeting affinity molecules, called Affibody molecules, were generated through phage selection. The aim of this thesis was to produce three corresponding affinity maturation libraries based on the three potential PSMA binders and select for improved binders through phage display. The affinity maturation libraries were produced, and two to four selection cycles were performed with yet insufficient enrichment. Therefore, more cycles must be performed in order to discover potential PSMA-binders with sufficient affinity.Prostataspecifikt membranantigen (PSMA) är ett membranprotein som uttrycks i både prostatacancerceller såväl som i nybildade blodkärl kring en tumör. Cancercellers uttryck av PSMA kan användas för att både behandla och upptäcka tumörer i kroppen. Tidigare har tre PSMA-sökande affinitetsmolekyler tagits fram, kallade Affibodymolekyler, genom fagdisplay. Syftet med detta examensarbete var att producera tre korresponderande affinitetsmatureringsbibliotek baserat på de tidigare potentiella PSMA-bindarna, och att selektera via fagdisplay för att hitta bindare med högre affinitet. De tre affinitetsmatureringsbiblioteken producerades och två till fyra selektionscykler har genomförts utan att se tillräcklig anrikning av bindare. Därför måste fler cykler genomföras i syfte att hitta potentiella PSMA-bindare med tillräckligt god affinitet

Inference of glioblastoma migration and proliferation rates using single time-point images

Cancer cell migration is a driving mechanism of invasion in solid malignant tumors. Anti-migratory treatments provide an alternative approach for managing disease progression. However, we currently lack scalable screening methods for identifying novel anti-migratory drugs. To this end, we develop a method that can estimate cell motility from single end-point images in vitro by estimating differences in the spatial distribution of cells and inferring proliferation and diffusion parameters using agent-based modeling and approximate Bayesian computation. To test the power of our method, we use it to investigate drug responses in a collection of 41 patient-derived glioblastoma cell cultures, identifying migration-associated pathways and drugs with potent anti-migratory effects. We validate our method and result in both in silico and in vitro using time-lapse imaging. Our proposed method applies to standard drug screen experiments, with no change needed, and emerges as a scalable approach to screen for anti-migratory drugs. The spatial positioning of cultured glioblastoma cells is used to estimate cell motility and drug effects from single end-point images in vitro