Polypeptide-DNA hydrogel and preparation method

A technology of hydrogel and polypeptide, which is applied in the field of biomedicine, can solve the problems that hydrogel needs to be deepened, and achieve the effect of good mechanical strength

Active Publication Date: 2014-07-09
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, preparing hydrogels with good biocompatibility and suitable mechanical strength remains a major challenge

Method used

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  • Polypeptide-DNA hydrogel and preparation method
  • Polypeptide-DNA hydrogel and preparation method
  • Polypeptide-DNA hydrogel and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1: Single-stranded DNA molecule (ssDNA) synthesis

[0065] Using the solid-phase phosphoramidite DNA synthesis method, the single-stranded DNA molecules shown in Table 1 were synthesized using an ABI394 DNA synthesizer, as follows:

[0066] Fix the solid-phase carrier containing a specific base in a DNA synthesizer, synthesize it according to the sequence shown in Table 1, and then use a concentrated ammonia solution for 3 hours at 60 degrees Celsius to ammonolyze it, remove ammonia through a vacuum concentrator, and pass the obtained crude product through a high-efficiency After purification by liquid chromatography (eluent: triethylamine acetate buffer, 100 mM, pH 7.0), the protective group containing DMT (dimethoxytrityl) was manually removed with 3% trifluoroacetic acid Then, the filter was washed with deionized water to remove DMT, and the target product was obtained. The structure of the target product was verified by MALDI-TOF-MS (Ionization Time-of-Flight ...

Embodiment 2

[0069] Example 2: Double-stranded DNA molecule (dsDNA) synthesis

[0070] According to the following steps, using the ssDNA obtained in Example 1 as a raw material, the double-stranded DNA molecules shown in Table 2 were synthesized, as follows:

[0071] Mix the stoichiometric ssDNA with 1×TBE buffer containing 100mM NaCl, then heat the resulting solution to 95 degrees Celsius, keep it warm for 3 minutes, and then cool it to room temperature within 2 hours to obtain the target double-stranded DNA molecule. Among them, 12a12b, RaRb, and HaHb have a cohesive end with a length of 12 bases, 8a8b has a cohesive end with a length of 8 bases, and 8Ma8Mb has a cohesive end with a length of 8 bases, but contains a mismatch site in its cohesive end point, RaRb and HaHb each independently have restriction endonuclease sites, respectively Ecor I site (5'-GAATTC-3') and BamH I site (5'-GGATCC-3').

[0072] Table 2

[0073] dsDNA

Embodiment 3

[0075] Embodiment 3: the preparation of polypeptide

[0076] Preparation of 5-azido-pentanoic acid

[0077] 5-Bromo-valeric acid (2.715 g, 15 mmol) was dissolved in 30 ml of dimethyl sulfoxide (DMSO) and heated to 40 °C, then sodium azide (3.25 g, 50 mmol) was gradually added, followed by heating at 85 °C , reacted overnight under stirring conditions, cooled the temperature of the obtained mixture to 40 degrees Celsius, gradually added concentrated hydrochloric acid (7mL), then stirred overnight, extracted the obtained reaction product with ether (5×80ml), collected the ether phase, It was washed successively with 10% sodium bicarbonate solution (2×100ml) and water (6×100ml), then dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a yellow oily product, namely 5-azido-valeric acid.

[0078] The analytical results of the yellow oily product are as follows:

[0079] 1 H-NMR (CDCl 3 ,400MHz):δ1.63-1.76(m,4H),2.40(t,J=7.4Hz,2H),3.30(t,J=6.9Hz,2H...

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Abstract

The invention provides a polypeptide-DNA hydrogel and a preparation method, wherein, the polypeptide-DNA hydrogel comprises the following components: polypeptide, wherein a single chain DNA molecule enables covalent binding on the polypeptide; a double chain DNA molecule, wherein the double chain DNA molecule has two cohesive ends; wherein the cohesive ends and the single chain DNA molecule are complementary, and polypeptide forms a crosslinking structure through the complementation of the cohesive ends of the double chain DNA molecule and the single chain DNA molecule. The mechanical strength of the polypeptide-DNA hydrogel can be adjusted, the biocompatibility is good, and the polypeptide-DNA hydrogel has reversible thermal responsiveness and specific enzyme responsiveness.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a polypolypeptide-DNA hydrogel and a preparation method thereof. Background technique [0002] Hydrogel is a kind of polymer containing hydrophilic groups, which can be swelled by water and maintain its complete three-dimensional network structure. It can absorb a large amount of water and swell significantly in water, and can continue to maintain its original structure without being dissolved after significant swelling. It can sense small changes in external stimuli, such as temperature, pH value, ionic strength, electric field, magnetic field, etc. , and can respond to external stimuli. Because the structure of hydrogel is very similar to the extracellular matrix that provides the biological environment for cells, hydrogel has broad application prospects in the field of biomedicine. However, preparing hydrogels with good biocompatibility and suitable mechanical strength...

Claims

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Application Information

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IPC IPC(8): C08J3/24C08J3/075C12N5/071
Inventor 刘冬生李志波李闯陈平杨忠强金娟邢永政
Owner TSINGHUA UNIV
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