Enrichment method of low abundance protein in bee venom

A protein, low abundance technology, applied in the field of protein processing, can solve the problems of protein loss, poor method reproducibility, cumbersome separation process, etc., and achieve high specificity, simple operation, and good repeatability.

Inactive Publication Date: 2013-07-17
BEE RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest disadvantage of these two methods as traditional protein separation and purification methods is that the separation process is cumbersome, and many proteins are lost during the extraction and purification process, resulting in protein loss, so that some proteins with specific biological functions have no

Method used

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  • Enrichment method of low abundance protein in bee venom

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0032] Example 1 Enrichment of low-abundance proteins in the venom of the bee bee in Italy

[0033] 1. Soak 500 mg of hexapeptide ligand beads bonded with porous polyhydroxymethacrylate particles in a syringe for 5 minutes in 2.0 mL of pure water, then mix on a shaker for 1 minute, centrifuge for 2 minutes, discard the effluent, and repeat the above Operate once.

[0034] 2. Add 2.0 mL PBS buffer solution to the above syringe, shake for 1 min, centrifuge for 2 min, discard the effluent, and repeat the above operation once.

[0035] 3. Accurately weigh 1.0g of the bee venom sample of Italian honeybees and place it in a 2.5mL centrifuge tube, add 1.0mL pure water, vortex to make it completely dissolved, then centrifuge at high speed at 4℃ and 10000rpm for 5min, and take the supernatant .

[0036] 4. Place the sample supernatant in the above syringe containing hexapeptide ligand beads and mix for 30 minutes, and mix in a shaker for 1 minute every 5 minutes to make the sample supernatant...

Example Embodiment

[0045] Example 2

[0046] 1. Soak 300 mg of hexapeptide ligand beads bonded with porous polyhydroxymethacrylate particles in a syringe with 1.0 mL of pure water for 5 minutes, then mix on a shaker for 1 minute, centrifuge for 2 minutes, discard the effluent, and repeat the above Operate once.

[0047] 2. Add 1.0 mL of PBS buffer solution to the above syringe, shake for 1 min, centrifuge for 2 min, discard the effluent, and repeat the above operation once.

[0048] 3. Accurately weigh 1.0g of the bee venom sample of Italian honeybees and place it in a 2.5mL centrifuge tube, add 1.0mL pure water, vortex to make it completely dissolved, then centrifuge at high speed at 4℃ and 10000rpm for 5min, and take the supernatant .

[0049] 4. Place the sample supernatant in the above syringe containing hexapeptide ligand beads and mix for 25 minutes, and mix for 1 minute in a shaker every 5 minutes to make the sample supernatant fully contact the beads, centrifuge for 2 minutes, and discard the f...

Example Embodiment

[0054] Example 3

[0055] 1. Soak 500 mg of hexapeptide ligand beads bonded with porous polyhydroxymethacrylate particles in a syringe with 2.5 mL of pure water for 5 minutes, then mix on a shaker for 3 minutes, centrifuge for 5 minutes, discard the effluent, and repeat the above Operate once.

[0056] 2. Add 2.0 mL PBS buffer solution to the above syringe, shake for 3 minutes, centrifuge for 5 minutes, discard the effluent, and repeat the above operation once.

[0057] 3. Accurately weigh 1.0g of the bee venom sample of Italian honeybees into a 2.5mL centrifuge tube, add 2.0mL pure water, vortex to completely dissolve it, then centrifuge at high speed at 4℃, 10000rpm for 5min, and take the supernatant .

[0058] 4. Place the sample supernatant in the above syringe containing hexapeptide ligand beads and mix for 40 minutes. Mix in a shaker for 1 minute every 5 minutes to fully contact the sample supernatant with the beads, centrifuge for 2 minutes, and discard the flow out liquid.

...

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Abstract

The invention relates to an enrichment method of low abundance protein in bee venom. The method includes: using hexapeptide ligand microspheres bonded with porous poly(2-hydroxyethyl methacrylate) granules as absorbent base, mixing the absorbent base with bee venom to absorb the protein in the bee venom, washing with PBS (phosphate buffered saline) buffer solution to remove excess high-abundance protein, and eluting the protein absorbed to the absorbent base with eluent to obtain the eluent with the enriched low-abundance protein. The enrichment method of low abundance protein is efficient, stable, well repeatable and highly specific and has great significance to enhancing the study on low-abundance protein, the search on information of more allergens in bee venom, the development of traditional Chinese medicines, the study on disease diagnosis markers and occurrence mechanisms of disease, and the like.

Description

technical field [0001] The invention relates to the field of protein treatment, in particular to a method for enriching low-abundance proteins in bee venom. Background technique [0002] The main bee species raised in our country is the Italian honeybee with strong production capacity, which has strong disease resistance and cold resistance. Bee venom is a liquid secreted by the venom glands and accessory glands of worker bees and stored in the poison sac. Italian bee venom has a variety of pharmacological and biological activities, mainly including anti-inflammatory, antihypertensive, analgesic, antiviral, etc., especially for the treatment of rheumatism, rheumatoid arthritis, frozen shoulder, and cardiovascular diseases , tumors and multiple sclerosis and other diseases have a good therapeutic effect (Jilin Science and Technology Press, 2000, 1-121). In order to fully understand the relevant information of all low-abundance proteins in bee venom, all proteins should be e...

Claims

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Application Information

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IPC IPC(8): C07K1/22
Inventor 周金慧李熠吴黎明赵静薛晓锋
Owner BEE RES INST CHINESE ACAD OF AGRI SCI
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