Protein container, polynucleotide, vector, expression cassette, cell, method for producing container, method for pathogen recognition or disease diagnosis, use of container, and diagnostic kit

A technology of protein and protein structure, applied in the application of pharmacy and biotechnology, falling into the field of chemistry, preparation, and medicine, can solve the problems of unproved simultaneous expression

Pending Publication Date: 2022-03-29
FUNDACAO OSWALDO CRUZ FIOCRUZ
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, other patent literatures have proposed GFP variants targeting the expression of peptides in protein loops, however, these studies have not demonstrated the simultaneous expression of more than 4 peptides in different insertion sites in GFP protein loops without losing any of their essential properties. Feasibility of features (WO02090535, US2003224412, WO200134824)

Method used

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  • Protein container, polynucleotide, vector, expression cassette, cell, method for producing container, method for pathogen recognition or disease diagnosis, use of container, and diagnostic kit
  • Protein container, polynucleotide, vector, expression cassette, cell, method for producing container, method for pathogen recognition or disease diagnosis, use of container, and diagnostic kit
  • Protein container, polynucleotide, vector, expression cassette, cell, method for producing container, method for pathogen recognition or disease diagnosis, use of container, and diagnostic kit

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no. 1 approach

[0124] In a first embodiment, the invention relates to a diagnostic kit comprising a protein container as defined above.

[0125] Finally, the invention is described in detail by the examples given hereinafter. It should be emphasized that the present invention is not limited to these embodiments, and it also includes changes and modifications within the range that can be developed. It is also worth noting that the authorized use of all biological sequences of the Brazilian genetic heritage is registered with SISGEN under registration number AC53976.

Embodiment 1

[0127] Example 1 -Container protein construction

[0128] Amino acid sequences between different instances of green fluorescent protein, eGFP (GenBank: L29345.1; UniProtKB-P42212), Cycle-3 (GenBank: CAH64883.1), SuperFolder (GenBank: AOH95453.1), Split (Cabantous et al. Reports 3:2854, 2013), Superfast (Fisher&DeLisa.PLoS One 3:e2351, 2008), used to construct the new protein of the present invention. Sequence alignment and comparison were performed by Intaglio software (Purgatory Design, V3.9.4). From these data, certain changes were made so that the container protein could achieve the desired properties.

[0129] Changes were made to create restriction enzyme sites. The insertion of these sites was designed such that the physicochemical properties of the GFP protein could not be altered, thus not affecting the properties or qualities described in this patent application. Furthermore, the insertion of these restriction sites will allow the genetic manipulation of these pr...

Embodiment 2

[0197] Example 2 – Construction of the PlatCruzi protein

[0198] The "platform" container protein was genetically engineered to possess T. cruzi epitopes, which we refer to herein as the PlatCruzi platform. The gene corresponding to the PlatCruzi protein, referred to herein as the PlatCruzi gene, is described in the nucleotide sequence of SEQ ID NO:17.

[0199] Considering experimental data on the specificity and sensitivity of a diagnostic test for Chagas disease, a polyamino acid sequence derived from Trypanosoma cruzi was selected from available prior art literature (Peralta JM et al. J Clin Microbiol 32:971- 974,1994; Houghton RL et al. J Infect Dis 179:1226-1234,1999; Thomas et al. Clin Exp Immunol 123:465-471,2001; Rabello et al., 1999; Gruber & Zingales, Exp Parasitology, 76(1):1-12,1993 ; Lafaille et al., Molecular Biochemistry Parasitology, 35(2):127-36, 1989). The 10 multiple amino acid sequences referred to herein as TcEp1 to TcEp10 were selected for insertion ...

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Abstract

The present invention relates to a protein container into which various exogenous polyamino acid sequences can be inserted simultaneously for expression in various systems and for various uses. The present invention relates to polynucleotides that can produce protein containers mentioned above. The invention also relates to vectors and expression cassettes comprising the above mentioned polynucleotides. The invention also relates to a cell comprising the vector or the expression cassette mentioned above. The invention also relates to methods for producing said protein container and identifying pathogens or diagnosing diseases in vitro. The invention also relates to the use of said protein container and a kit comprising said protein container for diagnosis or as a vaccine composition.

Description

technical field [0001] The present invention falls within the fields of application of chemistry, pharmacy, medicine, biotechnology, and more specifically, of preparations for biomedical purposes. The present invention relates to protein receptacles that can concomitantly accept multiple exogenous polyamino acid sequences for expression in various systems and for various uses. The present invention relates to polynucleotides capable of producing the above-mentioned protein containers. The invention also relates to vectors and expression cassettes comprising the aforementioned polynucleotide(s). The present invention further relates to cells comprising the aforementioned vector or expression cassette. The invention further relates to methods for producing said protein containers and for pathogen recognition or in vitro disease diagnosis. The invention further relates to the use of said protein container and a kit comprising said protein container for diagnostic purposes or a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K19/00A61K38/17A61K39/005A61K39/10A61K39/12A61K39/35A61K39/205A61K39/215C12N15/12C12N15/62C12N15/63G01N33/566G01N33/569
CPCA61K39/12G01N33/569Y02A50/30G01N33/582C12N15/70C12N2760/20134A61P31/14C12N2770/20034C07K14/005A61K2039/55566A61K39/099A61K39/35A61K39/005C07K16/10C07K14/43595A61K38/00C07K2319/60C07K2319/00
Inventor 小大卫·威廉·普罗旺斯安德烈莎·达马塔·杜兰斯帕洛玛·N·佩戈S·G·德西蒙
Owner FUNDACAO OSWALDO CRUZ FIOCRUZ
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