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Peptides capable of binding to titanium, silver and silicone

A technology of binding capacity and conjugates, which can be used in hybrid peptides, compound screening, peptides, etc., and can solve problems such as increased binding capacity

Inactive Publication Date: 2006-09-06
JAPAN SCI & TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As a result, it was found that the bonding ability with titanium was increased

Method used

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  • Peptides capable of binding to titanium, silver and silicone
  • Peptides capable of binding to titanium, silver and silicone
  • Peptides capable of binding to titanium, silver and silicone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Add 10 mg of titanium particles with a particle size of 150 μM (Sumitomo Chitanium, Hyogo) to a 1.5 ml eppendorph tube, and use 500 μl of 50 mM Tris (hydroxymethyl) methylamine (hereinafter, Tris (Kishida Chemical, Osaka)) pH 7.5 0.1% bovine serum albumin (hereinafter BSA) 0.1% polyoxyethylene sorbitan monolaurate (hereinafter, Tween- 20 (SIGMA Company, St.louis)) and washed twice. The titanium particles were precipitated by a desktop centrifuge H1300 (コクサン) at 13,000 rpm for 5 seconds, and the supernatant was removed to clean the titanium particles. After washing, in order to block the non-specific adsorption of the phage, rotator RT-50 (Taitech Co., Ltd.) was further used to rotate and stir in 1 ml of the same solution at room temperature for 30 minutes.

[0082] The titanium particles were precipitated by centrifugation in a desktop centrifuge H1300 (コクサン) at 13,000rpm for 5 seconds. After removing the supernatant, a phage library containing a peptide display was ad...

Embodiment 2

[0093] Using the phage clones obtained in Example 1, the binding ability to titanium was evaluated according to the following experiment. The same method as the panning operation shown in Example 1 was carried out. Different from Example 1, the stirring time between titanium and phage clones was 1 hour. When the concentration of Tween20 in each solution was 0.5%, and the concentration of BSA was 1%, the clone obtained from D12 according to the added phage titer was 10 9 pfu, 10 for clones obtained from C7C 11 pfu-implementation. The titanium-binding abilities of the individual phage clones are summarized in FIG. 5 .

Embodiment 3

[0095]According to Example 2, especially for the clone (presenting the peptide of SEQ ID NO: 3) that strongly binds to titanium, it is measured by the crystal oscillator-shaped biomolecular interaction analyzer QCM-D300 (q-sense AB company, イエテボリ) To study the binding mode of titanium surface and phage.

[0096] In the crystal oscillator, the titanium sensor of the QCM-D300 genuine product is used. The temperature is set at 24.99°C, and the measured value is from 24.68°C to around 24.70°C. Measured under the condition of blocking the sensor with BSA and under the condition of not blocking the sensor. When the sensor is not blocked with BSA, after measuring the reference value with TBS, the titer of the phage measured in sequence is adjusted to 10 10 pfu / ml of phage solution. When blocking with BSA, after measuring the reference value with TBS, incubate with TBS and 0.1% BSA for about ten minutes to block, wash the free BSA with TBS again, and adjust the titer of phage to 10...

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Abstract

The present invention provides a peptide sequence, a phage, an artificial protein or a chimeric molecule having a binding ability to titanium, silver, silicon, necessary to confer higher capacity of titanium, silver, silicon material with the use of soft matters, or to provide a complex of a peptide, a phage, an artificial protein or a chimeric molecule, and titanium, having the peptide sequence and functional peptide sequence. By bringing into contact a population of phage wherein said phage of said population collectively express a library of different peptide sequence, recovering titanium bound to phage particles via peptide sequence by centrifugation, proliferating the obtained phage particles in bacteria, and repeating panning operation and concentrating titanium binding phage clones. Among the phage clones, peptide RKLPDAPGMHTW and the like is identified. As for a peptide having a binding ability to titanium, silver, silicon, RKLPDA or RALPDA can be exemplified.

Description

technical field [0001] The present invention relates to a method for screening peptides with binding ability to titanium, peptides with binding ability to titanium, silver and / or silicon, peptides with binding ability to titanium, silver and / or silicon Titanium combined with titanium, silver or silicon , silver or silicon and peptide complexes, peptides with binding capabilities to titanium, silver and / or silicon, artificial proteins combined with functional peptides or functional proteins, such artificial proteins combined with titanium, silver or silicon, titanium, A complex of silver or silicon and an artificial protein, a combination of a peptide with the ability to bind to titanium, silver and / or silicon and a marker substance or a peptide tag, or a chimeric protein combined with a non-peptide compound, such a chimeric protein Complexes of titanium, silver or silicon and chimeric proteins bound to titanium, silver or silicon, phage surface displaying peptides with the abi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K7/08C07K19/00C12N1/00C12N7/00G01N33/53G01N33/553G01N33/569G01N33/84
CPCG01N33/5308C07K7/06C07K7/08G01N2500/00G01N33/56983G01N33/553G01N33/84
Inventor 芝清隆佐野健一
Owner JAPAN SCI & TECH CORP
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