Publications by authors named "Florian Tomszak"

CAR T cell therapy has been an effective treatment option for hematological malignancies. However, the therapeutic potential of CAR T cells can be reduced by several constraints, partly due to immunogenicity and toxicities. The lack of established workflows enabling thorough evaluation of new candidates, limits comprehensive CAR assessment.

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Pichia pastoris (syn. Komagataella phaffii) represents a commonly used expression system in the biotech industry. High clonal variation of transformants, however, typically results in a broad range of specific productivities for secreted proteins.

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Yeast surface display (YSD) has been shown to represent a powerful tool in the field of antibody discovery and engineering as well as for selection of high producer clones. However, YSD is predominantly applied in Saccharomyces cerevisiae, whereas expression of heterologous proteins is generally favored in the non-canonical yeast Pichia pastoris (Komagataella phaffii). Establishment of surface display in P.

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Antibody phage display is the most commonly used in vitro selection technology for the generation of human recombinant antibodies and has yielded thousands of useful antibodies for research, diagnostics, and therapy. The prerequisite for successful generation of antibodies using phage display is the construction of high-quality antibody gene libraries. Here, we give the detailed methods for the construction of human immune and naive scFv gene libraries.

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Since the development of therapeutic antibodies the demand of recombinant human antibodies is steadily increasing. Traditionally, therapeutic antibodies were generated by immunization of rat or mice, the generation of hybridoma clones, cloning of the antibody genes and subsequent humanization and engineering of the lead candidates. In the last few years, techniques were developed that use transgenic animals with a human antibody gene repertoire.

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Article Synopsis
  • Antibody phage display technology enables the generation of human antibodies necessary for diagnostics and therapy, and this study focuses on developing optimized libraries for better antibody selection.
  • Two naive antibody gene libraries, HAL9 and HAL10, were created from 98 healthy donors, achieving a theoretical diversity of 1.5×10(10) independent clones, with improved production of soluble antibodies through changes in tagging methods.
  • Analysis of selected antibodies showed that both the CDR-H3 length and amino acid diversity were critical, confirming that all CDR-H3 amino acids in the human repertoire can be effectively used in antibody generation without bias from the expression system.
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