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Method for preparing biology adhesive using emerald mussel adhesion protein

A mussel adhesion protein and bioadhesive technology, which is applied in surgical adhesives, applications, medical science, etc., can solve the problems of high price, difficulty in meeting needs, and few adhesion proteins, and achieve low cost and easy preparation. Convenience, Inexpensive Effects

Inactive Publication Date: 2009-06-10
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the amount of Adhesin secreted by blue mussels is very small, and only 1 mg of Adhesin can be extracted from about 10,000 mussels. Therefore, the Adhesin product CELL-TAK obtained from this TM The price is very expensive, it is difficult to meet the needs

Method used

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  • Method for preparing biology adhesive using emerald mussel adhesion protein
  • Method for preparing biology adhesive using emerald mussel adhesion protein
  • Method for preparing biology adhesive using emerald mussel adhesion protein

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Embodiment 1: the preparation of emerald mussel adhesive protein bioadhesive (Pvfp)

[0024] 1. Obtaining and homogenizing the feet of Emerald Mussels: Take live Emerald Mussels with a shell length of 13-14 cm, cut off their feet, weigh the feet with a fresh weight of 30 g, wash them in ice-precooled PBS, and put them in Contain 0.5% methanol pepstatin A (China Solarbio company) 30μM, leupeptin (Amersco company in the United States) 30μM and PMSF (BBI company in the United States) 1mM ice-cold 180ml 5% acetic acid, use T18 basic ( Germany IKA Company) tissue disperser, homogenate on ice for 2 minutes, pour the homogenate into a centrifuge tube, centrifuge at 20000g, 4°C for 60 minutes;

[0025] 2. Protein acidification: Discard the precipitate after centrifugation, take the supernatant S1, and record the volume V1, place the supernatant S1 on ice for pre-cooling, and slowly add 60% perchloric acid (PCA) to a final concentration of 1% (V 60％PCA =V1 / 59ml), stirred gently...

Embodiment 2

[0028] Example 2: Adhesion of Emerald mussel adhesion protein bioadhesive (Pvfp) on different material surfaces in a humid environment

[0029] Add 10 μl of emerald mussel bioadhesive with a concentration of 1.44 mg / ml dropwise on the cleaned plastic, glass, metal titanium and other materials, and add bovine serum albumin (BSA, bovine serum albumin) with the same concentration at the same time and CELL-TAK TM Make negative and positive controls respectively, place them in a humid environment at 25°C for 12 hours, take them out and dry them, wash them with distilled water for 2 hours, and then stain them with Coomassie Brilliant Blue R250, as figure 1 As shown, the results show that on the surface of glass slide a and titanium material b, the bioadhesive of emerald mussel adhesion protein (Pvfp) and CELL-TAK TM The adhesion ability of CELL-TAK is similar, while on the surface of plastic Petri dish c, Emerald mussel adhesion protein bioadhesive (Pvfp) has a higher than CELL-TAK...

Embodiment 3

[0030] Embodiment 3: Emerald mussel adhesive protein bioadhesive (Pvfp) bonding laboratory plastic consumables

[0031] Add 4 μl of 1.44 μg / μL emerald mussel bioadhesive dropwise on a laboratory consumable plastic Petri dish, place a white plastic pipette tip on the protein spot, and use BSA as a negative control. Place in a humid environment at 25°C for 12 hours. see results figure 2 , the emerald mussel adhesive protein bioadhesive (Pvfp) bonded the plastic Petri dish to the pipette tip together, which BSA failed to perform.

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Abstract

The invention provides a method for preparing biological adhesive by using perna viridis to adhere protein, relates to a biological adhesive. The invention provides a method for preparing a waterproof biological adhesive with better biological compatibility and low cost that extracts adhesive protein from perna viridis having wide raw material and low cost. The method includes steps as follows: cutting foot from live perna viridis, then mincing and homogenating, processing protein acidification, acetone protein precipitation, purification, finally, obtaining the waterproof biological adhesive. The adhesive protein waterproof biological adhesive can be obtained rapidly from perna viridis most abundant in China southeast seaside. The perna viridis biological adhesive has characteristics of high production, low cost and better adhesive effect. The prepared adhesive can adhere on surface of glass, metallic titanium and plastic, adheres cell in short time, adheres plastic or mouse thighbone, and has no cell toxicity. The biological adhesive as biological medical material biological adhesive has wide application prospect in clinical use.

Description

technical field [0001] The invention relates to a bioadhesive, in particular to a method for preparing a waterproof bioadhesive by extracting adhesive proteins from emerald mussels. Background technique [0002] Marine mussels belong to the mollusk Phylumbranchia, and are a kind of organisms commonly found in coastal and offshore areas. Their silk glands can secrete silk to attach to the hard substrate, so that they can still adhere tightly under the washing of huge waves. on the reef without separation. The main component of this silk viscose, the silk adhesive protein, has high strength, high toughness and water resistance. At present, BD Bioscience in the United States has extracted adhesion proteins from the feet of blue mussels (Mytilus edulis), which are used as cell and tissue adhesion agents. The product has been commercialized, and the name and article number are respectively BD CELL-TAK TM CELL AND TISSUE ADHESIVE, Catalog No. 354240, 354241. This product can b...

Claims

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Application Information

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IPC IPC(8): A61L24/10
Inventor 张其清金利华刘加鹏蒋臻
Owner XIAMEN UNIV
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