Unlock instant, AI-driven research and patent intelligence for your innovation.

Separation purification process for heterogenous homogeneous serum albumin

A whey protein, separation and purification technology, applied in the field of separation and purification of heterologous and homologous whey proteins, can solve the problems of difficulty in separation, difficulty in amplification, and similar physical and chemical properties, and achieves improved recovery rate, stable production process, and product purity. high effect

Inactive Publication Date: 2009-02-18
INST OF PROCESS ENG CHINESE ACAD OF SCI
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In addition to the human whey protein expressed by the transgene, there is also the whey protein expressed by the mammal itself in the milk of transgenic mammals. The heterologous and homologous whey proteins have similar structures, close physical and chemical properties, and are difficult to separate. Wang Jianwu (recombinant human a- Identification and analysis of whey albumin transgenic cattle, doctoral thesis of China Agricultural University, 2007) reported that bovine and human a-lactalbumin were separated by anion exchange isocratic elution, but it was difficult to effectively scale up

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Separation purification process for heterogenous homogeneous serum albumin
  • Separation purification process for heterogenous homogeneous serum albumin
  • Separation purification process for heterogenous homogeneous serum albumin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Take 100mL of transgenic human-derived a-lactalbumin milk, the total protein concentration of bovine milk is 33g / L, the concentration of human-derived a-lactalbumin is 1.4g / L, and the concentration of bovine-derived a-lactalbumin is 1.0g / L. Sigma3k30 centrifuge at 4 degrees Celsius, adjust the centrifugal force to 10,000g and centrifuge for 30 minutes to remove lipids and casein, then collect the supernatant to obtain 80mL of whey, the total protein concentration is 9.5g / L, and the concentration of human a-whey albumin is 1.7g / L, the concentration of bovine a-lactalbumin is 1.2g / L. Use 1M hydrochloric acid or sodium hydroxide solution to adjust the pH of the whey to 8.0, slowly add ammonium sulfate powder until the whey ammonium sulfate saturation is 45%, and the molar count is 2.09M. After shaking at room temperature for 2 hours, the precipitate Samples were centrifuged in a Sigma3k30 centrifuge at 4°C with a centrifugal force of 10,000g for 30 minutes and 70mL of the...

Embodiment 2

[0032]Take 100mL of transgenic human-derived a-lactalbumin milk, the total protein concentration of bovine milk is 33g / L, the concentration of human-derived a-lactalbumin is 1.4g / L, and the concentration of bovine-derived a-lactalbumin is 1.0g / L. Sigma3k30 centrifuge at 4 degrees Celsius, adjust the centrifugal force to 10,000g and centrifuge for 30 minutes to remove lipids and casein, then collect the supernatant to obtain 80mL of whey, the total protein concentration is 9.5g / L, and the concentration of human a-whey albumin is 1.7g / L, the concentration of bovine a-lactalbumin is 1.2g / L. Use 1M hydrochloric acid or sodium hydroxide solution to adjust the pH of the whey to 7.0, slowly add ammonium sulfate powder until the whey ammonium sulfate saturation is 40%, and the molar count is 1.84M. After shaking at room temperature for 2 hours, the precipitate The sample was centrifuged at 4 degrees Celsius in a Sigma 3k30 centrifuge with a centrifugal force of 10,000g for 30 minutes...

Embodiment 3

[0035] Take 100mL of transgenic human-derived a-lactalbumin milk, the total protein concentration of bovine milk is 33g / L, the concentration of human-derived a-lactalbumin is 1.4g / L, and the concentration of bovine-derived a-lactalbumin is 1.0g / L. Sigma 3k30 centrifuge at 4 degrees Celsius, adjust the centrifugal force to 10000g and centrifuge for 30 minutes to remove lipids and casein, then collect the supernatant to obtain 80mL of whey, the total protein concentration is 9.5g / L, and the concentration of human a-whey albumin is 1.7 g / L, the concentration of bovine a-lactalbumin is 1.2g / L. Use 1M hydrochloric acid or sodium hydroxide solution to adjust the whey pH to 7.0, use Shanghai Zhixin Instrument Co., Ltd. DDB-300 constant current peristaltic pump and Millipore company 100KD hollow fiber membrane column for ultrafiltration, cycle 5 times, and circulate buffer Use 20mM, pH7.0 phosphate buffer, finally collect 80mL of permeated liquid, the total protein concentration of th...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention relates to a method for separating and purifying heterogenous and homogeneous whey proteins. The method includes the following steps: genetically modified mammal milk containing the heterogenous and homogeneous whey proteins is chosen as material; whey is obtained after lipid and casein are removed by centrifugation; the obtained whey is preprocessed, removing macromolecular impurities such as immune globulin; the whey or the preprocessed whey undergoes hydrophobic interaction chromatography separation; the heterogenous and homogeneous whey proteins in the whey are interacted with hydrophobic chromatography media; because the spacial conformations and hydrophobicity of the heterogenous and homogeneous whey proteins are different, the heterogenous and homogeneous whey proteins can be separated in hydrophobic chromatography; and further, the pure products of the heterogenous and homogeneous whey proteins with high purity can be obtained by gel permeation chromatography. The method has the advantages of high recovery rate and stable production technique and is suitable for mass production.

Description

technical field [0001] The invention belongs to a method for separating and purifying proteins, in particular to a method for separating and purifying heterologous and homologous whey proteins. Background technique [0002] Many whey proteins in mammalian milk contain high nutritional value and medicinal value, such as providing various essential amino acids for the body, inhibiting bacteria and inducing tumor cell apoptosis. There are allergens in mammalian milk such as whey protein in bovine milk, which can cause IgE or non-IgE-mediated allergic reactions in some infants and adults. Using transgenic technology to express transgenic human whey protein in mammalian mammary gland reactor can improve the nutritional value of emulsion, and human whey protein purified from transgenic emulsion as raw material can be used as a food additive with high nutritional value and high economic added value. [0003] In addition to the human whey protein expressed by the transgene, there i...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/20C07K1/16
Inventor 苏志国张焱罗坚闭静秀马光辉卫强
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI