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Frank Breitling
Prof. Dr. Frank Breitling
Peptide arrays and antibody libraries

Phone: +49 721 608-23859
frank breitlingNrc3∂kit edu

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Peptide Arrays and Antibody Libraries

Prof. Dr. Frank Breitling

Our research is segmented in two different research areas: (1) The combinatorial synthesis of high-density peptide arrays with the help of solid amino acid particles (in cooperation with our spin-off company PEPperPRINT) and (2) the generation of a library of >105 different human hybridomas (more) that should speed up considerably the generation of monoclonal antibodies.

High-density peptide arrays



In addition to the xerographic methods (laser printer or CMOS chip with pixel electrode arrays), the combinatorial patterning of the amino acids can be realized with the direct transfer of amino acids due to the laser ablation from the donor slides to the synthesis support (Fig. 5) [8]. With this cLIFT (combinatorial Light Induced Forward Transfer) method, we synthesized, for instance, HA- (Tyr-Pro-Tyr-Asp-Val-Pro-Asp-Tyr-Ala) and Flag-peptides (Tyr-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) with a spot pitch of 150 µm. Hereby, activated OPfp amino acids were deposited and coupled to the surface in repetitive manner.  Fig. 6 shows the results of the specific fluorescent staining. The significant advantage of the cLIFT-method is the possibility of coupling non-activated amino acids [9]. If two layers with a monomer and an activation reagent are deposited on each other, the molecules can mix with each other by heating the substrate due to the nanometer thickness of those layers. Thus, a non-inactivated amino acid can be deposited upon the activator, and dozens of thousands chemical reactions can starts simultaneously by heating the substrate. We believe that the method of the “stacked nanolayers” is applicable to a wide class of materials and chemical reactions.

The long term goal of this research is a procedure to synthesize freely chosen peptides in good quality on a glass slide, and for a few Euros chemical costs. Thereby, and for the first time, completely new applications for peptide arrays should be feasible. (more)









Fig. 5; (a-h) The combinatorial laser induced transfer of synthesis building blocks. Donor-slide has different synthesis building blocks embedded into polymer matrix (labeled with different colors in (b) and (c)). The donor slides are positioned on the acceptor slide (a) with the transfer material downwards. A laser transfers small amounts of the material onto the acceptor slide. Patterns with different monomers are generated with laser transfer with different donor slides. The coupling of the monomers (e) begins with the heating of the acceptor slide. Subsequently, the unreacted monomers are washed away (f) and the usual steps for the peptide chemistry, such as acetylation of unreacted amino groups and the removal of the protective groups, are carried out (g). The repetition of the cycle (a-g) leads to the combinatorial peptide synthesis in the array format (h). (I) cLIFT synthesizer. The laser radiation is modulated with an acousto-optic modulator (AOM) and directed to the scanning system. Donor and acceptor slides are automatically exchanged by a slide loader.


Fig. 6; Fluorescence images of the cLIFT-synthesized patterns of Flag and HA peptides stained with specific anti-Flag and anti-HA antibodies. Scale: (a) 2 mm, (b) 1 mm, (c) 250 μm.









[8] Loeffler F F, Foertsch T C, Popov R, Mattes D S, Schlageter M, Sedlmayr M, Ridder B, Dang F-X, von Bojničić-Kninski C, Weber LK, Fischer A,Greifenstein J, Bykovskaya V, Buliev I, Bischoff F R, Hahn L, Meier M A R, Bräse S, Powell A K, BalabanT S, Breitling F, Nesterov-Mueller A. High-flexibility combinatorial peptide synthesis with laser-based transfer of monomers in matrix material. Nature Communications, 2016, DOI: 10.1038/NCOMMS11844

[9] Loeffler F F, Breitling F, Nesterov-Mueller A, Kombinatorische Chemie im hochdichten Arrayformat, BIOspectrum, 2016, 5: 532-534; DOI:10.1007/s12268-016-0720-1