Biology subjects

Ziegler, M. J.

Publications and source records attributed to Ziegler, M. J..

3 recordsLinked to original sources

bioRxiv2025

Discovery and optimization of a guanylhydrazone-based small molecule to replace bFGF for cell culture applications

Replacing growth factors with a synthetic alternative molecule is an attractive opportunity to increase consistency, scalability and cost-effectiveness of cell-based products. Herein, we describe the discovery of a chemical class of FGFR1 agonists mimicking the action of basic fibroblast growth factor (bFGF), an essential component of cell-culture media. The guanylhydrazone-based molecule, TCB-32, was identified via structure-based virtual screening on the orthosteric binding site of FGFR1. It was shown to significantly increase cell proliferation by activating the FGFR1 signalling pathway like bFGF and exhibited enhanced thermostability over bFGF by containing activity over the course of several days. After extensive structure-activity relationship studies it was possible to increase potency and efficacy leading to three highly potent agonists. This finding has the potential to remove current bottlenecks in large-scale cell production as required for applications like cultivated meat or cell therapy. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/635029v1_ufig1.gif" ALT="Figure 1"> View larger version (49K): org.highwire.dtl.DTLVardef@1d734aaorg.highwire.dtl.DTLVardef@1c28e0corg.highwire.dtl.DTLVardef@1fbdaa2org.highwire.dtl.DTLVardef@1cedd5e_HPS_FORMAT_FIGEXP M_FIG C_FIG HIGHLIGHTSO_LIDiscovery of a highly potent small molecule to replace bFGF in serum-free medium C_LIO_LIActivation of proliferation through the FGFR1-signalling pathway C_LIO_LIEnhanced thermal stability over bFGF C_LIO_LISAR led to three small molecules with decreased EC50, named TCB-494, TCB-541 and TCB-621 C_LI

biochemistry↗

bioRxiv2020

Bioorthogonal red and far-red fluorogenic probes for wash-free live-cell and super-resolution microscopy

Small-molecule fluorophores enable the observation of biomolecules in their native context with fluorescence microscopy. Specific labelling via bioorthogonal tetrazine chemistry confers minimal label size and rapid labelling kinetics. At the same time, fluorogenic tetrazine-dye conjugates exhibit efficient quenching of dyes prior to target binding. However, live-cell compatible long-wavelength fluorophores with strong fluorogenicity have been difficult to realize. Here, we report close proximity tetrazine-dye conjugates with minimal distance between tetrazine and fluorophore. Two synthetic routes give access to a series of cell permeable and impermeable dyes including highly fluorogenic far-red emitting derivatives with electron exchange as dominant excited state quenching mechanism. We demonstrate their potential for live-cell imaging in combination with unnatural amino acids, wash-free multi-colour and super-resolution STED and SOFI imaging. These dyes pave the way for advanced fluorescence imaging of biomolecules with minimal label size.

cell biology↗

bioRxiv2020

A chemical strategy to control protein networks in vivo

Direct control of protein interaction by chemically induced protein proximity (CIPP) holds great potential for cell- and synthetic biology as well as therapeutic applications. However, toxicity, low cell-permeability and lack of orthogonality currently limit the use of available chemical inducers of proximity (CIP). We present Mandi, a novel CIP and demonstrate its applicability in cell culture systems as well as living organisms for protein translocation, protein network shuttling and manipulation of endogenous proteins.

synthetic biology↗