Meguro-ku, Japan
Meguro-ku, Japan

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Ishizawa T.,University of Tokyo | Kawakami T.,University of Tokyo | Reid P.C.,PeptiDream Inc. | Murakami H.,University of Tokyo
Journal of the American Chemical Society | Year: 2013

Here, we describe a novel method that enables high-speed in vitro selection of functional peptides, peptidomimetics, and proteins via a simple procedure. We first developed a new cell-free translation system, the TRAP system (transcription-translation coupled with association of puromycin linker), which automatically produces a polypeptide library through a series of sequential reactions: transcription, association of puromycin-DNA linker, translation, and conjugation between the nascent polypeptide and puromycin-DNA linker. We then applied the TRAP system for the selection of macrocyclic peptides against human serum albumin. Six rounds of selection using TRAP display were performed in approximately 14 h, yielding macrocyclic peptides with nanomolar affinity to their target protein. Because TRAP display enables high-speed selection of functional polypeptides, it will facilitate the generation of various polypeptides that are useful for biological and therapeutic applications. © 2013 American Chemical Society.


Patent
University of Tokyo and Peptidream Inc. | Date: 2014-08-04

An object of the present invention is to provide a method of producing a peptide containing a charged non-proteinogenic amino acid in a cell-free translation system, and the like. The present invention provides a method of producing a peptide containing a charged non-proteinogenic amino acid. It includes a step of expressing a peptide in a cell-free translation system including (i) at least one tRNA to which a non-proteinogenic amino acid having a protecting-group-introduced charged group has been bound and (ii) a nucleic acid that encodes the peptide and contains at least one codon corresponding to an anticodon of the tRNA; and a step of removing the protecting group of the non-proteinogenic amino acid residue contained in the peptide.


Patent
University of Tokyo and PeptiDream Inc. | Date: 2016-06-15

An object of the present invention is to provide a method of producing a peptide containing a charged non-proteinogenic amino acid in a cell-free translation system, and the like. The present invention provides a method of producing a peptide containing a charged non-proteinogenic amino acid. It includes a step of expressing a peptide in a cell-free translation system including (i) at least one tRNA to which a non-proteinogenic amino acid having a protecting-group-introduced charged group has been bound and (ii) a nucleic acid that encodes the peptide and contains at least one codon corresponding to an anticodon of the tRNA; and a step of removing the protecting group of the non-proteinogenic amino acid residue contained in the peptide.


Patent
University of Tokyo and Peptidream Inc. | Date: 2013-09-02

An object of the present invention is to provide a VEGFR2 inhibitor peptide having high specificity and available at a low cost. The present invention provides a peptide having the following amino acid sequence: [wherein, Xaa_(2 )represents Val or derivative thereof, Xaa_(6 )represents Asp or derivative thereof, Xaa_(7 )represents Pro or derivative thereof, Xaa_(8 )represents Trp or derivative thereof, Xaa_(10 )represents Asn or derivative thereof, Xaa_(11 )represents Gly or derivative thereof, Xaa_(12 )represents Leu or derivative thereof, Xaa_(1), Xaa_(3 )to Xaa_(5), Xaa_(9), and Xaa_(13 )to Xaa_(15 )each independently represent an arbitrary amino acid or derivative thereof]; or [wherein, Xaa_(24 )represents His or derivative thereof, Xaa_(25 )represents Pro or derivative thereof, and Xaa_(16 )to Xaa_(23 )and Xaa_(26 )to Xaa_(30 )each represent an arbitrary amino acid].


Patent
University of Tokyo and Peptidream Inc. | Date: 2014-05-12

An object of the present invention is to provide a method for producing a peptide library capable of incorporating an arbitrary number of arbitrary proteinogenic and/or non-proteinogenic amino acids in an arbitrary site. The invention provides a method for producing a peptide library including 110^(6 )or more kinds of peptides containing amino acids encoded by N_(1)N_(2)N_(3), including a step of preparing an mRNA library including mRNAs which encode peptides of the peptide library and each contain at least one N_(1)N_(2)N_(3); and a step of translating each mRNA of the mRNA library in a cell-free translation system added with tRNA containing an anticodon to any one of N_(1)N_(2)N_(3 )codons and charged with an amino acid corresponding to the codon (wherein, N_(1), N_(2), and N_(3 )are each independently selected from adenine (A), guanine (G), cytosine (C), and uracil (U) and an arbitrary amino acid is reassigned to each N_(1)N_(2)N_(3)).


Patent
University of Tokyo and PeptiDream Inc. | Date: 2016-03-16

An object of the present invention is to provide a method for producing a peptide library capable of incorporating an arbitrary number of arbitrary proteinogenic and/or non-proteinogenic amino acids in an arbitrary site. The invention provides a method for producing a peptide library including 110^(6) or more kinds of peptides containing amino acids encoded by N_(1)N_(2)N_(3), including a step of preparing an mRNA library including mRNAs which encode peptides of the peptide library and each contain at least one N_(1)N_(2)N_(3); and a step of translating each mRNA of the mRNA library in a cell-free translation system added with tRNA containing an anticodon to any one of N_(1)N_(2)N_(3) codons and charged with an amino acid corresponding to the codon (wherein, N_(1), N_(2), and N_(3) are each independently selected from adenine (A), guanine (G), cytosine (C), and uracil (U) and an arbitrary amino acid is reassigned to each N_(1)N_(2)N_(3)).


Patent
University of Tokyo and PeptiDream Inc. | Date: 2015-07-15

An object of the present invention is to provide a VEGFR2 inhibitor peptide having high specificity and available at a low cost. The present invention provides a peptide having the following amino acid sequence:_(2) represents Val or derivative thereof, Xaa_(6) represents Asp or derivative thereof, Xaa_(7) represents Pro or derivative thereof, Xaa_(8) represents Trp or derivative thereof, Xaa_(10) represents Asn or derivative thereof, Xaa_(11) represents Gly or derivative thereof, Xaa_(12) represents Leu or derivative thereof, Xaa_(1), Xaa_(3) to Xaa_(5), Xaa_(9), and Xaa_(13) to Xaa_(15) each independently represent an arbitrary amino acid or derivative thereof]; or_(24) represents His or derivative thereof, Xaa_(25) represents Pro or derivative thereof, and Xaa_(16) to Xaa_(23) and Xaa_(26) to Xaa_(30) each represent an arbitrary amino acid].


Patent
PeptiDream Inc. | Date: 2012-08-29

Provided are linkers suitable for preparing a conjugate of a nucleic acid and a peptide as a translation product thereof in a reconstituted cell-free translation system in genotype-phenotype mapping (display methods), said linkers comprising a single-stranded structure region having a side chain base pairing with the base at the 3-end of an mRNA at one end and a peptidyl acceptor region containing an amino acid attached to an oligo RNA consisting of a nucleotide sequence of ACCA via an ester bond at the other end, characterized in that the ester bond is formed by using an artificial RNA catalyst. Also provided are display methods using [mRNA] - [linker] - [peptide] conjugates assembled via such linkers.


Patent
PeptiDream Inc. | Date: 2015-12-09

An improved method used in selecting a useful protein, peptide, peptide analog by an evolution molecule engineering is provided. A transcription-linker association-translation coupling reaction system characterized by incorporation of a template DNA library to enable a step of forming translation product/linker/mRNA complexes through transcription of a template DNA library to mRNAs, association of mRNAs with linkers, translation of mRNAs, and binding with translation products to be automatically performed in a reaction system, comprising factors necessary for transcription, factors necessary for translation, and linkers.


Patent
Peptidream Inc. | Date: 2014-01-29

An improved method used in selecting a useful protein, peptide, peptide analog by an evolution molecule engineering is provided. A transcription-linker association-translation coupling reaction system characterized by incorporation of a template DNA library to enable a step of forming translation product/linker/mRNA complexes through transcription of a template DNA library to mRNAs, association of mRNAs with linkers, translation of mRNAs, and binding with translation products to be automatically performed in a reaction system, comprising factors necessary for transcription, factors necessary for translation, and linkers.

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