Cow milk protein fatty acid modifier capable of inducing human breast cancer cell apoptosis, and preparation method thereof

A technology of human breast cancer cells and breast cancer cells, which is applied in the field of milk protein fatty acid modification and its preparation for inducing apoptosis of human breast cancer cells, can solve the problems of limited sources and difficult large-scale preparation, and promote intensive processing Effect

Inactive Publication Date: 2015-03-11
GUANGDONG OCEAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, since human lactalbumin mainly exists in human breast milk, the

Method used

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  • Cow milk protein fatty acid modifier capable of inducing human breast cancer cell apoptosis, and preparation method thereof
  • Cow milk protein fatty acid modifier capable of inducing human breast cancer cell apoptosis, and preparation method thereof
  • Cow milk protein fatty acid modifier capable of inducing human breast cancer cell apoptosis, and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0029] In the greenhouse, dissolve lactalbumin in 0.05mM EDTA, 10mM Tris-HCl, pH8.5 solution, overnight at 0~4℃ to remove Ca 2+ , concentrated by a protein ultrafiltration tube to obtain decalcified α-lactalbumin in an unfolded state; the final protein concentration is 100 μM; in a greenhouse, oleic acid is mixed with absolute ethanol, and ultrasonically fully dissolved to make a fatty acid stock solution; use Take the stock solution before adding 10mM Tris-HCl (pH8.5), the final concentration of ethanol in the solution is not more than 2% (V / V), the final concentration of oleic acid is 500μM; add the above fatty acid solution to DEAE-Sepharose fast flow anion column, using NaCl gradient elution to disperse fatty acids; add the above lactalbumin solution to an anion column pretreated with fatty acids; and use low ion concentration gradient (0.2M NaCl, 10mM Tris, pH8.5) Unbound fatty acids and milk proteins are then eluted with high-salt buffer (1M NaCl, 10mM Tris, pH8.5), and ...

Embodiment 2

[0031] In the greenhouse, dissolve lactalbumin in 1mmol / L EDTA, 20mM Tris-HCl, pH8.0 solution, overnight at 0~4℃ to remove Ca 2+, concentrated by protein ultrafiltration tubes to obtain decalcified α-lactalbumin in an unfolded state; the final protein concentration is 50 μM; in the greenhouse, oleic acid is mixed with absolute ethanol, and ultrasonically fully dissolved to make a fatty acid stock solution; use Take the stock solution before adding 20mM Tris-HCl (pH8.0), the final concentration of ethanol in the solution is not more than 2% (V / V), and the concentration of oleic acid is 800μM; add the above fatty acid solution to the DEAE-Sepharose fast flow anion column , using NaCl gradient elution to disperse the fatty acid; add the above lactalbumin solution to the anion column pretreated with fatty acid; and use low ion concentration gradient (0.2M NaCl, 10mM Tris, pH8.0) The combined fatty acid and milk protein are then eluted with high-salt buffer (1M NaCl, 10mM Tris, pH8...

Embodiment 3

[0033] In the greenhouse, dissolve lactalbumin in 0.08mM EDTA, 15mM Tris-HCl, pH8.2 solution, overnight at 0~4℃ to remove Ca 2+ , concentrated by a protein ultrafiltration tube to obtain decalcified α-lactalbumin in an unfolded state; the final protein concentration is 120 μM; in a greenhouse, linoleic acid is mixed into absolute ethanol, and ultrasonically fully dissolved to make a fatty acid stock solution; Take the stock solution before use and add 15mM Tris-HCl (pH8.2), the final concentration of ethanol in the solution is not more than 2% (V / V), the final concentration of linoleic acid is 1800μM; add the above fatty acid solution to DEAE-Sepharose fast flow anion column, using NaCl gradient elution to disperse fatty acids; add the above lactalbumin solution to an anion column pretreated with fatty acids; and use low ion concentration gradient (0.2M NaCl, 10mM Tris, pH8.5) to elute Unbound fatty acids and milk proteins were removed respectively, and then eluted with high-s...

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Abstract

The present invention relates to a cow milk protein fatty acid modifier capable of inducing human breast cancer cell apoptosis, and a preparation method thereof, and belongs to the technical field of medicine or function food preparation. According to the method, cow milk albumin is subjected to a decalcification treatment to form the unfolded state, and then an unsaturated fatty acid is adopted to carry out appropriate molecule modification on the lactoalbumin component through the anion exchange chromatography technology to form the milk protein fatty acid modifier, wherein the in vitro cell model test results of the modifier prove that the modifier can significantly induce the human breast cancer cell (including estrogen receptor positive cells and estrogen receptor negative cells) apoptosis and does not significantly affect the normal human breast cells. According to the present invention, the support can be provided for development of the breast cancer prevention and treatment drug and the anti-breast cancer function food, the new reference is provided for the high value utilization of the whey protein and the optimization of the formula milk powder, and the positive significance is provided for promotion of the fine and deep processing of the milk product.

Description

technical field [0001] The invention relates to a fatty acid modification of milk protein and a preparation method thereof, in particular to a fatty acid modification of milk protein capable of inducing apoptosis of human breast cancer cells and a preparation method thereof. Belonging to the technical field of medicine or functional food preparation, the milk protein fatty acid modification prepared by the invention can induce the apoptosis of human breast cancer cells without significant influence on normal human breast cells. Background technique [0002] Cancer has become the number one killer that endangers human health and has become one of the largest public health problems in the world. The incidence of cancer is increasing year by year. It is estimated that by 2020, there will be 16 million new cancer patients and 10 million people will die every year. in cancer (International Union Against Cancer UICC, 2011). [0003] Although countries around the world have invest...

Claims

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Application Information

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IPC IPC(8): A61K38/40A61P35/00
Inventor 钟赛意刘寿春谌素华王维民秦小明
Owner GUANGDONG OCEAN UNIVERSITY
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