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Artificial multimeric proteins with programmable biological and chemical functions
Poster Title: Artificial multimeric proteins with programmable biological and chemical functions
Submitted on 27 Oct 2017
Author(s): Piotr M. Skowron, Łukasz Janus, Olga Żołnierkiewicz, Małgorzata Skowron, Joanna Jeżewska-Frąckowiak, Daria Krefft, Dawid Nidzworski, Kasjan Szemiako, Natalia Maciejewska, Marta Nowak, Alicja Wiercińska-Drapało, Grzegorz Węgrzyn, Aneta Szymańska, Agnieszka Żylicz-Stachula
Affiliations: University of Gdansk, Faculty of Chemistry, Dept. of Molecular Biotechnology, Gdansk, Poland
This poster was presented at Advances in Analytical & Bio-analytical Chemistry
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Poster Information
Abstract: Artificial multimeric proteins with programmable biological and chemical functions

The need for novel biomaterials is rapidly increasing due to progress and fusion of molecular biology, chemistry and nanotechnology. Some of these are constructed with the use of genetic engineering methods and the obtained DNA constructs are cloned and expressed in microorganisms. Resulting recombinant enzymes and other proteins are useful in scientific research as well as in medical diagnostics, forensic and biological analyses, as well as in the biotechnology industry. Generation of recombinant biomaterials often involves nucleic acids amplification using the Polymerase Chain Reaction. We present an alternative amplification scheme, adding defined sections of DNA to the nascent concatameric DNA molecule combined with an expression vector, to produce recombinant, artificial, concatameric proteins and RNAs. Monomeric DNA sections, used as precursors for amplification, can have a built-in coded function, for example an immunogenic epitope. The method allows the construction of DNA concatamers ordered in a head-to-tail-orientation. A very high number of polymerized DNA segments, at least 500 copies, can be combined into a single DNA molecule in an ordered fashion. Once amplified, the concatameric artificial gene, present in an amplification-expression vector, directs the transcription of concatameric RNA from fused vector promoters, and leads to the formation of a concatameric protein with a pre-programmed function. These functions/applications include: construction of vaccines with a highly increased potential for stimulating the immune system; combination of different protein epitopes/domains into a single artificial protein in one step, as coded on the designed synthetic DNA monomer; polyproteins containing modules for rare metals recovery from sea water; toxic metal- or organic and inorganic compounds binding for environmental remediation; detoxification after poisoning; new generation biosimilar drugs for regenerative medicine; protective concatameric polypeptides with monomers based on activators or inhibitors of biological functions for the treatment of molecular and infectious diseases.

Acknowledgments: this project was supported by the National Center for Research and Development, Warsaw, Poland, grant no. STRATEGMED1/235077/9/NCBR/2014 and POIG.01.04.00-22-140/12; Jagiellonian Center for Innovation, Cracow, Poland; SATUS VC, Warsaw, Poland and BioVentures Institute Ltd, Poznan, Poland.
Summary: We present an alternative to PCR amplification combined with an expression vector, to produce recombinant, concatameric proteins and RNAs. Monomeric DNA sections, used as precursors for amplification, can have a built-in coded function. Expressed concatameric protein have a pre-programmed functions, such as vaccines, hybrid protein epitopes/domains, rare metals recovery, toxic metal and compounds binding for environmental remediation, body detoxification, biosimilar drugs.References: 1. Skowron P, Zylicz-Stachula A, Zolnierkiewicz O, Skowron M, Janus L, Jezewska-Frackowiak J, Krefft D, Nidzworski D, Szemiako K, Maciejewska N, Nowak M, Szymanska A: A method of obtaining a polyepitopic protein as well as a DNA vector for embodying this method. WO Patent Application 2015162560 A1 (2015).
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