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Biohybrid Nanoarchitectures:
Protein Adaptor Based Nano-Object Assembly
(PABNOA)

Functional Expansion of the Cell:
De Novo Organells &
Cellular Modules and Nanoreactors

Biogenic Materials:
Elastine like Proteins
ECM mimicking matrix proteins

Sustainable Biopolymers &
Scaffolds in
Regenerative Medicine

Bioorthogonal Chemistry & Bioconjugation:
Unnatural Amino Acids
Glycolipid Library Tool Box

Modular Molecular Tecton Library
for Biogenic Materials
by One-Vector-Toolbox-Platform (OVTP)

Bioprocess & Enzyme engineering
Peptide Generator
for Biopharmaceuticals

Biomimetic Systems
Membranebiochemistry for
tethered bilayer lipid membranes (tBLM)




       










































Concept and method for the efficient design and production of biogenic materials


OVTP 01



The OVTP allows for sequential-directional ligation (SDL) and cyclic-ligation (CL) of DNA-tectons


CL & SDL
Sequential-directional ligation (SDL) and Cyclic-Ligation (CL) for multimerisation and defined arrangement of DNA-tectons
I. Schematic illustration of the SDL process (on the left). The DNA-tectons prepared from the identical OVTP vectors by sequential action of the restriction enzymes SacI and EarI (donor). By the sequential action of the restriction enzymes BspQI and SacI the same plasmid is used as an
acceptor for compatible DNA-tectons.

II. In order to generate repetitive protein multimers a CL reaction was performed by alternating reaction temperatures with alternating enzymatic activities as a "one tube reaction" (on the right). OVTP vectors with different DNA-tectons were used as donors and acceptors for the preparation and insertion of other DNA-tectons. Tectons were added serial to each cycle as an uniform fraction or parallel as a mixed fraction at each cycle to generate homogeneous or randomly configurated complex DNA-tectons.
 
Combinations of defined preparations of OVTP vectors in combination with fitting DNA-tectons allows for the precise composition of DNA-tectons
within the OVTP vector.
Resulting vectors could subsequently serve as acceptor or donor vector for further combinations on DNA-level or for protein-tecton expression.
CL & SDL 02
Subsequent generation of DNA-tecton and protein-tecton libraries constituted from various building blocks


tecton library 01tecton librar 02




Huber, M. C., Schreiber, A., Wild, W., Benz, K., Schiller, S. M. "Introducing a combinatorial DNA-toolbox platform constituting defined protein-based biohybrid materials" Biomaterials 2014, 35, 8767-8779

Schiller, Stefan M., "Protein Tectons in Synthetic Biology:  The expansion of cellular functionality combining chemical biology of small organic molecules with protein tectons - unnatural amino acids, protein based biohybird materials & de novo organelles", in Synthetic Biology, ed. Giese, B., von Gleich, A., Pade, C., Wigger, H. (Springer, 2014, ISBN 978-3-319-02782-1)

Schiller, S., Huber, M. C. "Protein Assembler" 2013 PCT/EP2013/060957, WO 2013/178627

                                                      



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Revised April 2015 by M. C. Huber