Patents

2010 to 2021 (selected patents)

Breitling F, Bykovskaya V, Leibe K, Löffler F, Märkle F, Nesterov-Müller A, Schillo S, von Bojnicic-Kninski C. (2014) Method for combinatorial particle manipulation for producing high-density molecule arrays, in particular peptide arrays, and molecule arrays that can be obtained by means thereof. Patent family EP2986370B1, US9925509B2, JP6502319B2. Licensed to SME PEPperPRINT.

Topic: Multi-material nano3D printer. (more)
Similar to the method that is described above, chemically activated amino acid building blocks are embedded into a solid polymer, but this time formulated into a material layer on top of a light absorbing polyimide foil (= “donor slide”). Instead of printing particles, a 2D laser scanning system transfers tiny material spots to selected areas of an acceptor. These material spots are nano-dimensioned in z-direction, which allows us to stack many different materials in freely chosen combinations (“nanostacks”). (more) 
The machine can be used to synthesize very high-density peptide arrays, (more) but also in many other types of extremely miniaturized chemical reactions. (more)

Althuon DS, Breitling F, Bykovskaya V, Loeffler F, Nesterov-Mueller A, Popov R, Ridder B, Von Bojnicic-Kninski C. (2016) Ultrahigh-density oligomer arrays and method of production thereof. Patent family EP3362463B1, DE102015117567B4, US10376858B2. Held by KIT.

Topic: One-cavity-one-peptide-method. (more)
This patent family adds another trick to solid materials-based synthesis of peptide arrays. Similar to the method that is described above, chemically activated amino acid building blocks are embedded into a solid polymer and formulated into monodisperse particles. Instead of printing these particles, they are randomly used to fill >2,5 Mio cavities on a glass slide that are slightly larger than the particles. In other words: each cavity picks exactly one particle. There, particles are melted to elongate many different peptides on the array at once. Particles are colour-coded, e.g. green fluorescence means “Fmoc-Alanine-OPfP ester”. This allows us to easily find out for each and every cavity the sequence of the synthesized peptides, by doing a fluorescence scan for every synthesis layer.

1999-2009 (selected patents)

Breitling F, Poustka A, Groß KH, Dübel S, and Saffrich R. (1999). Method and devices for applying substances to a support, especially monomers for the combinatorial synthesis of molecule libraries. Patent family AU773048B2, EP1140977B1, US9752985B2, JP4471329B1. Licensed to PEPperPRINT GmbH.

Topic: Solid-material-based chemical synthesis. (more)
This patent family covered solid materials-based synthesis of peptide arrays. Briefly, chemically activated amino acid building blocks are embedded into a solid polymer and formulated into particles. Either a laser printer or a computer chip is used to print these particles to designated areas on an acceptor. There, particles are melted to elongate many different peptides on the array at once.

Breitling F, Moldenhauer G, Lüttgau S, Kühlwein T, and Poustka A. (2001) Methods of producing protein libraries and selection of proteins from them. Patent family EP1298207B1, WO2003029458A2.

Breitling F, Moldenhauer G, and Poustka A. (2000). Selection of monoclonal antibodies. Patent family EP1141271B1, WO2000042176A1. Licensed to SME Abeome.

Topic: Easy humanisation of mouse monoclonal antibodies. Surface displayed antibodies on eukaryotic cells.

Before 1999 (selected patents)

Breitling F, Fuchs P, Little M, and Dübel S. (1991) Recombinant antibodies at the surface of E.coli. Patent family DE4122598, EP547200B1, US5591604. Licensed to Bayer AG.

Topic: Libraries of recombinant antibodies. Surface displayed antibodies on bacteria.

Breitling F, Little M, Dübel S, Braunagel M, and Klewinghaus I. (1991) Phagemid for antibody screening. Patent family EP547201B1, EP1065271B1, JP6500930A, US6730483B2, US6387627B1. Licensed to Behringwerke and to SME Affimed.

Breitling F, Little M, Dübel S, Seehaus T, and Klewinghaus I. (1990) Herstellung und Verwendung von Genbanken synthetischer menschlicher Antikörper ("synthetic human-antibody-libraries"), Patent family AU1991070115B2, EP440146B1, US5840479, JP4211395, IE75220B1. Licensed to Behringwerke and to SME Affimed.

Breitling, F., Dübel, S., Little, M., Seehaus, T., and Klewinghaus, I. (1990) Preparation and use of gene banks of human antibodies (human antibody libraries). Patents DE4002898, EP440147B1 (issued 2004), US6319690, JP3514778B1 (issued 2004). Status: Licensed to Bayer AG.

Topic: Libraries of recombinant antibodies. In parallel with patents from Cambridge (Dr. Greg Winter), these patents covered all successful commercial methods to generate libraries of recombinant antibodies.

Textbook

Rekombinante Antikörper; Lehrbuch und Kompendium für Studium und Praxis. Zweite vollkommen neu überarbeitete Auflage
Dübel, S., Breitling, F., Frenzel, A., Jostock, Th., Marschall, A.L.J., Schirrmann, Th., Hust, M.
2019. Springer Spektrum; ISBN 978-3-662-50275-4

Recombinant Antibodies
Dübel, S.; Breitling, F.
1999. Wiley-Spektrum; ISBN 978-0471178477

Rekombinante Antikörper; Erste Auflage
Dübel, S.; Breitling, F.
1997. Spektrum Akademischer Verlag; ISBN 978-3-8274-0150-2

Awards

Breitling F and Bischoff FR. 2^nd winner of the business plan competition Genius Biotech Award (March 2001; 40.000 DM, organised by the state Baden-Württemberg).

Breitling F, Bischoff FR, Wallich R, Poustka A, Stadler V, and Breitling F. Winner of Innovationswettbewerb Medizintechnik with the project Borrelia peptidome arrays (November 2001)

Stadler V, Bischoff FR, Felgenhauer T, Kring M, and Breitling F. 1^st winner of the business plan competition Science4Life with the project Fertigung und Vertrieb von Biochips zur Parallel-Synthese unterschiedlicher Peptide (June 2009, 30.000 €, nation-wide competition).

Breitling F, Bischoff FR, Stadler V, Felgenhauer T, Leibe K, Fernandez S (all from DKFZ or KIT) and Güttler S, Gröning M, Willems P, Biesinger B (Fraunhofer IPA). Winner of Wissenschaftspreis des Stifterverbandes with the project Peptide laser printer (May 2008, 50.000 €).

Before 2009 (selected publications)

Beyer, M.^*, Nesterov, A.^*, Block, I., König, K., Felgenhauer, T., Fernandez, S., Leibe, K., Torralba, G., Hausmann, M., Trunk, U., Lindenstruth, V., Bischoff, F.R.^*, Stadler, V.^*, and Breitling, F.^* (2007) Combinatorial synthesis of peptide arrays onto a computer chip’s surface. Science 318, 1888
^* shared first / last author

Stadler, V.^*, Felgenhauer, T.^*, Beyer, M., Fernandez, S., Leibe, K., Güttler, S., Gröning, M., Torralba, G., Hausmann, M., Lindenstruth, V., Nesterov, A., Block, I., Pipkorn, R., Poustka, A., Bischoff, F.R.^*, and Breitling, F.^* (2008) Combinatorial synthesis of peptide arrays with a laser printer. Angewandte Chemie International Edition. 47, 7132 –7135
^* shared first / last author

Topic: Solid-material-based chemical synthesis. (more)
These are the first papers that describe solid material-based, miniaturized & parallelized chemical synthesis. Briefly, chemical building blocks are embedded into a solid polymer and formulated into particles. Either a laser printer or a computer chip is used to print these particles to designated areas on an acceptor. There, particles are melted to start many different coupling reactions at once.
 

Breitling, F., Dübel, S., Seehaus, T., Klewinghaus, I., and Little, M. (1991) A surface expression vector for antibody screening. Gene 104, 147-153

Fuchs, P., Breitling, F., Dübel, S., Seehaus, T., and Little, M. (1991) Targeting recombinant antibodies to the surface of Escherichia coli: Fusion to a peptidoglycan associated lipoprotein. (Nature)Biotechnology 9, 1369-1372

Rondot, S., Anthony, K., Dübel, S., Ida, N., Wiemann, S., Beyreuther, K., Frost, L., Little, M., and Breitling, F. (1998) Epitopes fused to F-Pilin are Incorporated into Functional Recombinant Pili. Journal of Molecular Biology 279, 589-603

Rondot, S.^*, Koch, J.^*, Breitling, F., and Dübel, S. (2001) A helper phage to improve single-chain antibody presentation in phage display. NatureBiotechnology 19, 75-78
^* shared first author

Topic: Libraries of recombinant antibodies.
The Gene and (Nature)Biotechnology papers are the second and third papers to describe recombinant antibodies. The gene paper is the first paper to show that phagemids instead of normal filamentous phages can be used to display recombinant antibodies on a phage via a fusion protein (= antibody-pIII). All recombinant antibody libraries that are used today use this principle. The reason is simple: only when the phagemid – that codes for the recombinant antibody-pIII fusion protein – is packaged into particles, the expression of the antibody is turned on, resulting in only moderate selective pressure against productive antibody expression.

The Journal of Molecular Biology & Nature Biotechnology papers describe another important trick to build huge libraries of antibodies fused to phagemid particles (that code for the antibody): Using a wildtype phage with wildtype pIII envelope protein to package phagemids into particles means that most particles display only a few antibody-pIII proteins on their surface. Therefore, we designed a mutant helper phage that is devoid of wildtype pIII, and, therefore, is forced to use antibody-pIII when assembling the phage particle.