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Recombinant elastin and production thereof

A fluorescent protein and protease cleavage technology, which is applied in the field of recombinant elastin and its production, can solve problems such as difficult extraction of elastin

Pending Publication Date: 2022-01-28
GELTOR INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Extraction of elastin from animal tissue is difficult due to extensive crosslinking of elastin peptides

Method used

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  • Recombinant elastin and production thereof
  • Recombinant elastin and production thereof
  • Recombinant elastin and production thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 125

[0125] Example 1.25kDa truncated human elastin

[0126] Non-naturally occurring truncated human elastin without a His-tag, linker and / or thrombin cleavage site is disclosed below. The non-naturally occurring truncated human elastin is truncated relative to full length human elastin (SEQ ID NO: 20). The codon-optimized nucleotide and amino acid sequences encoding this elastin are disclosed below. In SEQ ID NO:4, the DsbA secretion tag is encoded by nucleotides 1-57 and encodes amino acids 1-19 of SEQ ID NO:5. In SEQ ID NO:4, the non-naturally occurring truncated human elastin sequence is encoded by nucleotides 58-927 and encodes amino acids 20-309 of SEQ ID NO:5.

[0127] The codon-optimized nucleotide sequence encoding the non-naturally occurring truncated elastin is provided in SEQ ID NO:4.

[0128] ATGAAAAAGATTTGGCTGGCGCTGGCTGGTTTAGTTTTAGCGTTTAGCGCATCGGCGGGTCCGCAACCTGGGGTTCCGTTAGGTTATCCGATTAAAGCACCGAAACTGCCCGGCGGTTATGGTCTGCCGTACACAACCGGTAAACTGCCGTATGGTTATGGCCCGGGTGGAGTT...

Embodiment 3

[0160] Example 3. 60kDa truncated human elastin

[0161] The amino acid sequence of a 60.3 kDa non-naturally occurring truncated human elastin internally truncated relative to full-length human elastin (SEQ ID NO:20) is disclosed in SEQ ID NO:10. The 60.3 kDa non-naturally occurring truncated human elastin has amino acids 461-527 deleted from full length human elastin (SEQ ID NO: 20).

[0162] GGVPGAIPGGVPGGVFYPGAGLGALGGGALGPGGKPLKPVPGGLAGAGLGAGLGAFPAVTFPGALVPGGVADAAAAYKAAKAGAGLGGVPGVGGLGVSAGAVVPQPGAGVKPGKVPGVGLPGVYPGGVLPGARFPGVGVLPGVPTGAGVKPKAPGVGGAFAGIPGVGPFGGPQPGVPLGYPIKAPKLPGGYGLPYTTGKLPYGYGPGGVAGAAGKAGYPTGTGVGPQAAAAAAAKAAAKFGAGAAGVLPGVGGAGVPGVPGAIPGIGGIAGVGTPAAAAAAAAAAKAAKYGAAAGLVPGGPGFGPGVVGVPGAGVPGVGVPGAGIPVVPGAGIPGAAVPGVVSPEAAAKAAAKAAKYGARPGVGVGGIPTYGVGAGGFPGFGVGVGGIPGVAGVPGVGGVPGVGGVPGVGISPEAQAAAAAKAAKYGAAGAGVLGGLVPGPQAAVPGVPGTGGVPGVGTPAAAAKSAAKVAAKAQLRAAAGLGAGIPGLGVGVGVPGLGVGAGVPGLGVGAGVPGFGAGADEGVRRSLSPELREGDPSSSQHLPSTPSSPRVPGALAAAKAAKYGAAVPGVLGGLGALGGVGIPGGVVGAGPA...

Embodiment 4

[0166] Example 4. Effect of Truncated Elastin on Fibroblast Viability, Tropocollagen Synthesis and Tropoelastin Synthesis ring

[0167] Human fibroblast cultures were used to assess the ability of the non-naturally occurring truncated human elastin molecules of Example 1 to affect tropoelastin and tropoelastin synthesis. Human fibroblast cultures were also used to determine the viability of human fibroblasts after exposure to truncated elastin.

[0168] A 2% w / w stock solution of truncated elastin was prepared from the truncated elastin of Example 1. Aliquots from the 2% truncated elastin stock solution were then used in the experiments described below.

[0169] Preparation of fibroblasts

[0170] Seed fibroblasts in 0.5 mL of fibroblast growth medium (FGM) in each well of a 24-well plate and incubate at 37 ± 2 °C and 5 ± 1% CO 2 Incubate overnight. The next day, remove the medium by aspiration to eliminate any non-adherent cells and replace with 0.5 mL of fresh FGM. ...

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Abstract

This disclosure provides non-naturally occurring truncated elastin molecules. The non-naturally occurring truncated elastin may improve the firmness, elasticity, brightness, hydration, tactile texture, and / or visual texture of skin. The non-naturally occurring truncated elastin may reduce degradation of the extracellular matrix.

Description

[0001] cross reference [0002] This application claims the benefit of U.S. Provisional Patent Application Nos. 62 / 833,415, filed April 12, 2019, and 62 / 889,397, filed August 20, 2019, which are hereby incorporated by reference in their entirety. Background technique [0003] Elastin is an elastic protein that is critical to the proper function of arteries, lungs, tendons, ligaments, skin and other tissues. Elastin provides tissue with the ability to stretch and return to its original shape. The protein tropoelastin is a building block of elastin. Unlike other structural proteins, such as collagen, which contain an array of genes, there is one gene for tropoelastin in humans. When the gene encoding tropoelastin is expressed, individual tropoelastin genes are spliced ​​to produce different forms of tropoelastin. Multiple tropoelastin molecules bind together to form elastin. [0004] The structure of natural elastin is similar to that of collagen, with an amino acid sequenc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78A61P17/00A61P17/16A61K38/00
CPCC07K14/78A61P17/16A61P17/00A61K38/00A61K8/64A61Q19/00A61Q19/08C07K2319/01C07K2319/21C07K2319/50C07K2319/60C12N15/70
Inventor 尼古拉·奥佐诺夫塔尼娅·伊莱亚森
Owner GELTOR INC
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