Dr. Humenik, MartinOpen Resume
Self-assembly is based on autonomous mechanism of non-covalent interactions between distinct building blocks. Related processes do not require external energy source and have no limitation in dimensional scaling. Such “bottom up” formation structures at different hierarchical levels is inspired by natural processes, which enable highly organized materials with exceptions properties such as bones, shells or spider silks.
Application of the self-assembly principles to control nano- and micro scaled structuring of biomaterials is in focus of our group.
Dr. Humenik, Martin
Nucleic acids could self-assemble into many different nano-structures with precision and predictability, which is unrivaled among the natural and synthetic materials. One of the best example for such high fidelity is represented by the DNA origami programming and folding. On the other hand, recombinant fibrous proteins also self-assemble into nanofibrils, supramolecular structures of densely packed cross-ß sheets. Such fibrils, typically 10 nm in diameter and hundreds of nanometers long, possess high physico-chemical stability as well as mechanical rigidity. Such systems represent attractive scaffolds for the generation of ordered nanomaterials.
To take advantage of both protein and DNA materials, we chemically combine recombinant spider silk proteins and nucleic acids in hybrid entities. Their fundamental properties are investigated to realize their utilization in materials research. One focus is the study of new chemical modifications and conjugations of the building blocks, arrangements protein moieties upon hybridizations of DNA, corresponding morphology and structure of conjugate fibrils. Further, we use specific DNA hybridization to trigger nucleation of the fibrils into hierarchical structures on surfaces. We combine soft- and photolithography techniques with the DNA hybrids to spatially define self-assembly of the supramolecular structures in 2D and 3D. Our technology enables formation of fibrillar nanohydrogel-like networks within arbitrary shaped microstructructures. Employing chemically coupled DNA aptameric functionalities, we can bind enzymes, growth factors or even whole cells specifically.
Fig. 1: Self-organization of DNA-protein hybrid materials in bottom up manner in solutions: Spider silk moieties in the hybrids, prepared by a chemical conjugation with DNA, were spatially arranged into branched structures using designed DNA hybridization and self-assembled into ribbons, which further self-organized into micro-rafts due to DNA interaction at specific temperature gradients.
Fig. 2: Pattering of the hybrid materials using DNA-directed hybridization technology: Surfaces modified by capture DNA were specifically linked to complementary DNA-spider silk conjugates on defined position using micro-contact printing technology. Immobilized conjugates served as nucleation sides for silk fibrils growth from the surface.
Fig. 3: Microstructured nanohydrogels. a) photolithography processed positive-tone photoresist with micro-wells for spatially defined protein self-assembly on the surface; b) AFM scans of the self-assembled fibrous microstructures after photoresist removal; c) immobilized fibrous networks reveal nano hydrogel properties like swelling and softening on the surface; d) cells specifically immobilized on the DNA-modified spider silk microstructures via pre-defined DNA-cell interaction.
Taken together, processing or structuring spider silk proteins and bio-functionalized hybrids thereof into nanohydrogel-based platform allows integrating different functionaries into immobilized fibril networks. Exploiting such biomolecular tools, more complex patterns consisting of multiple, unique functionalities, such as conjugated enzymes, aptamers or gold nanoparticles, deposited in 1D along the growing fibrils and in 2D pattern on surfaces are likely feasible. Concomitant possibility for specific immobilization of living cells as well as programmable nanohydrogel patterning offer a platform with great potential for development of numerous biological and biomedical applications in regenerative medicine and cell separations and analytics.
Fig. 4: Universal platform of spider silk nanofibrillar scaffold for multifunctional surface modifications
Kocourkova K., Musilova L., Smolka P., Mracek A., Humenik M., Minarik A.
Factors determining self-assembly of hyaluronan
Carbohydr. Polym. 254, 117307
Herold H.M., Döbl A., Wohlrab S., Humenik M., Scheibel T.
Designed Spider Silk-Based Drug Carrier for Redox- or pH-Triggered Drug Release
Biomacromolecules 21, 4904-4912
Wang Y., Stanzel M., Gumbrecht W., Humenik M., Sprinzl M.
Esterase 2-oligodeoxynucleotide conjugates as sensitive reporter for electrochemical detection of nucleic acid hybridization
Biosens.Bioelectron. 22, 1798-1806
Humenik M., Huang Y., Wang Y., Sprinzl M.
C-terminal incorporation of bio-orthogonal azide groups into a protein and preparation of protein-oligodeoxynucleotide conjugates by Cu(l)-catalyzed cycloaddition
ChemBioChem 8, 1103-1106
Humenik M., Poehlmann C., Wang Y., Sprinzl M.
Enhancement of Electrochemical Signal on Gold Electrodes by Polyvalent Esterase-Dendrimer Clusters
Bioconjug.Chem. 19, 2456-2461
Minarik A., Humenik M., Li S., Huang Y., Krausch G., Sprinzl M.
Ligand-Directed Immobilization of Proteins through an Esterase 2 Fusion Tag Studied by Atomic Force Microscopy
ChemBioChem 9, 124-130
Poehlmann C., Humenik M., Sprinzl M.
Detection of bacterial 16S rRNA using multivalent dendrimer-reporter enzyme conjugates
Biosens.Bioelectron. 24, 3383-3386
Poehlmann C., Wang Y., Humenik M., Heidenreich B., Gareis M., Sprinzl M.
Rapid, specific and sensitive electrochemical detection of foodborne bacteria
Biosens. Bioelectron. 24, 2766-2771
Koeck, K. S., Salehi, S., Humenik, M. & Scheibel, T.
Processing of Continuous Non-Crosslinked Collagen Fibers for Microtissue Formation at the Muscle-Tendon Interface
Advanced functional materials, 2021
Laomeephol, C., Vasuratna, A., Ratanavaraporn, J., Kanokpanont, S., Luckanagul, J., Humenik, M., Scheibel, Th. & Damrongsakkul, S.
Impacts of Blended Bombyx mori Silk Fibroin and Recombinant Spider Silk Fibroin Hydrogels on Cell Growth
Polymers. 2021, 13, 4182
Humenik, M., Winkler, A. & Scheibel, T.
Patterning of protein-based materials
Biopolymers, 2020; online Dec. 2020
Mathilde Lefevre, Patrick Flammang , A. Sesilja Aranko , Markus B. Linder , Thomas Scheibel , Martin Humenik , Maxime Leclercq , Mathieu Surin , Lionel Tafforeau, Ruddy Wattiez, Philippe Leclère, Elise Hennebert
Sea star-inspired recombinant adhesive proteins self-assemble and adsorb on surfaces in aqueous environments to form cytocompatible coatings
Humenik M., Preiß T., Goedrich S., Papastavrou G., Scheibel T.
Functionalized DNA spider silk nanohydrogels for controlled protein binding and release
Materials Today Bio, 6, 100045
Humenik M., Pawar K., Scheibel T.
Nanostructured, self-assembled spider silk materials for biomedical applications
In: S. Perrett et al. (eds.), Biological and Bio-inspired Nanomaterials, (Advances in Experimental Medicine and Biology 1174), 187-221
DeSimone E., Aigner T. B., Humenik M., Lang G., Scheibel T.
Aqueous electrospinning of recombinant spider silk proteins
Mater. Sci. Eng. C, 106, 110145
Humenik M., Scheibel T., Smith A.
Spider Silk: Understanding the Structure–Function Relationship of a Natural Fiber
Prog. Mol. Biol. Transl. Sci. 103, 131-185.
Humenik M., Smith A., Scheibel T.
Recombinant spider silks – biopolymers with potential for future applications
Polymers 3, 640–661
Humenik M., Scheibel T.
Self-assembly of nucleic acids, silk and hybrid materials thereof
J. Phys. Condens. Matter 26, 503102
Humenik M., Scheibel T.
Nanomaterial building blocks based on spider silk–oligonucleotide conjugates
ACS Nano 8, 1342-1349
Humenik M., Magdeburg M., Scheibel T.
Influence of repeat numbers on self-assembly rates of repetitive recombinant spider silk proteins
J. Struct. Biol., 186, 431-437
Humenik M., Markus D., Scheibel T.
Controlled hierarchical assembly ofspider silk-DNA chimeras into ribbons and raft-like morphologies
Nano Lett.., 14, 3999−4004
Humenik M., Smith A., Arndt S., Scheibel T.
Ion and seed dependent fibril assembly of a spidroin core domain
J. Struct. Biol. 191: 130–138
Humenik M., Smith A M., Arndt S., Scheibel T.
Data for ion and seed dependent fibril assembly of a spidroin core domain
Data in Brief 4: 571–576
Humenik M., Lang G., Scheibel T.
Silk nanofibril self-assembly versus electrospinning
Wiley Interdiscip. Rev.: Nanomed. Nanobiotechnol., 10: e1509
Molina A., Humenik M, Scheibel T.
Nanoscale patterning of surfaces via DNA directed spider silk assembly
Biomacromol., 20, 347-352
Humenik M., Mohrand M., Scheibel T.
Self-assembly of spider silk-fusion proteins comprising enzymatic and fluorescence activity.
Bioconjugate Chem., 29: 898 – 904.
Hardy J. G., Bertin A., Torres‐Rendon J. G., Leal‐Egaña A., Humenik M., Bauer, F., Walther A., Cölfen H., Schlaad H., Scheibel T.
Facile photochemical modification of silk protein-based biomaterials
Macromol. Biosci., 28: 1800216