High-throughput generation of bispecific binding proteins by sortase a-mediated coupling for direct functional screening in cell culture

High-throughput construction of multivalent binders and subsequent screening for biological activity represent a fundamental challenge: A linear increase of monovalent components translates to the square of possible bivalent combinations. Even high-efficiency cloning and expression methods become limiting when thousands of bispecific binders need to be screened for activity. In this study, we present an in vitro method for the efficient production of flexibly linked bispecific binding agents from individually expressed and purified monovalent binders. We established a sortase A-mediated coupling reaction to generate bispecific designed ankyrin repeat proteins (DARPins), with an optimized reaction maximizing the bivalent coupling product with low levels of monovalent side-products. These one-pot reaction mixtures could be used directly, without further purification, in cell-based assays. We generated a matrix of 441 different bispecific DARPins against the extracellular domains of the cancer-associated receptors EGFR, ErbB2, ErbB3, ErbB4, EpCAM, and c-MET and screened on two different ErbB2-positive cancer cells lines for growth-inhibitory effects. We identified not only known but also novel biologically active biparatopic DARPins. Furthermore, we found that the cancer cell lines respond in a highly reproducible and defined manner to the treatment with the 441 different bivalent binding agents. The generated response profiles can thus be used for functional characterization of cell lines because they are strongly related to the cell line-specific surface receptor landscape. Thus, our method not only represents a robust tool for screening and lead identification of bispecific binding agents, but additionally offers an orthogonal approach for the functional characterization of cancer cell lines.</p

Der Qualitätspakt E-Learning im Hochschulpakt 2020

Der vorliegende Tagungsband beinhaltet die Artikel zu den Vorträgen auf der GML² 2014 und eine Sammlung der zu der Tagung eingereichten Abstracts. Wir danken allen Referierenden für ihre anregenden Vorträge auf der Tagung und ihre Artikel für den Tagungsband, allen Vortragenden in den Workshops der Pre- Conference für ihre Beiträge und allen Einreichenden für das Zur-Verfügung- Stellen ihrer Abstracts für den Tagungsband

Empowering Diverse Learners through Design-Based Data Literacy Education: the Data Design Cycle Framework

Data literacy is a vital life-skill that is becoming increasingly necessary in today's society. As such, public data literacy education must become a substantive component in teaching, requiring an appropriate pedagogical model. This paper aims to present a new definition for data literacy, as well as conceptual framework for public data literacy education, drawing from tenets of design-based education and synthesized principles of data literacy education from literature. The resulting framework is termed the Data Design Cycle, and emphasizes collaboration, communication, and the development of problem-solving skills. The model is highly adaptable to different educational environments and learning styles, making it easily integrable into a variety of educational settings, including traditional classroom-based learning, online learning, and experiential learning programs. Furthermore, by promoting the principles of public teaching of data literacy and a learner-centred approach, the Data Design Cycle ensures that data literacy education is accessible to heterogeneous learners from diverse educational and socioeconomical settings

Informatics – Science of Minimal Systems with Maximal Complexity, In: Weert, Tom van (Eds.) ; Munro, Robert (Eds

It is a fundamental idea of computer science to search for, define, analyze, and operate with construction kits consisting of small sets of basic building blocks and a small number of operations to combine the building blocks to larger objects. While the construction kit is mostly simple, it often defines a vast and very complex field that consists of all possible objects that can be built from the building blocks by using any (finite) sequence of combinations of operators. This idea affects and structures many areas of computer science. We present examples from several fields, among them are imperative and functional programming languages, computable functions, Turing and register machines, Boolean functions, data types, object-oriented programming, characterizations of formal languages along with examples from other disciplines. How can informatics lessons profit from this observation? On the one hand, if lessons are oriented towards a fundamental idea, the idea may explain, structure, and integrate many different informatics subjects and phenomena by a single recurring scheme. On the other hand, since an idea like the construction kit principle also belongs to the sphere of everyday thinking, students already have a basic intuition of the concept which may enhance their understanding when entering any of the fields where the idea applies.

Philosophical Aspects of Fundamental Ideas: Ideas and Concepts

Abstract: We consider the term ªidea from a philosophical point of view. In particular we are interested in the concept s origins, its relevance to human thinking and in particular in its pedagogical value for computer science lessons in schools as well as universities. Since the concept of fundamental ideas in computer science has been seized, extended, and reviewed by other authors and applied to lessons, often with a different understanding of the defined terms, we wish to explain some of the objectives of the approach in more detail and in particular clarify the relation between concept and idea in order to provide a common understanding of the relevant notions.

Lernorte der Second-Life-Repräsentanz E-Learning 3D – didaktischer Einsatz und Nutzung für Großgruppen

Heeren J. Lernorte der Second-Life-Repräsentanz E-Learning 3D – didaktischer Einsatz und Nutzung für Großgruppen. In: Andreas S, Nicolas A, eds. Lernen im Digitalen Zeitalter. Dokumentation der Pre-Conference zur DeLFI2009 – Die 7. E-Learning Fachtagung Informatik der Gesellschaft für Informatik e.V. Potsdam/Berlin; 2009: 253-261