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Method for purifying lactoferrin from milk serum by using an expanded bed adsorption technology

A lactoferrin, expanding bed technology, applied to transferrin, animal/human proteins, specific peptides, etc., can solve the problems of cumbersome operation, long time, poor adsorption selectivity of lactoferrin, etc., and achieve simplified process and simple steps. The effect of easy operation and saving equipment investment

Inactive Publication Date: 2013-01-30
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since the FastLine SALTO medium has cation exchange and hydrophobic groups at the same time, the adsorption selectivity of lactoferrin is poor, and the protein components in milk are complex, and immunoglobulin and bovine serum albumin can be adsorbed, so more complicated washing is required. Lactoferrin with a purity greater than 90% can be obtained only by taking off steps, and the operation is relatively cumbersome and time-consuming

Method used

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  • Method for purifying lactoferrin from milk serum by using an expanded bed adsorption technology
  • Method for purifying lactoferrin from milk serum by using an expanded bed adsorption technology
  • Method for purifying lactoferrin from milk serum by using an expanded bed adsorption technology

Examples

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Effect test

Embodiment 1

[0035] 1) Add water to dissolve sweet whey powder to make 1 L of whey solution with a mass percent concentration of 50 g / L, and adjust the pH to 7.0 with NaOH;

[0036] 2) The expanded bed is filled with cation exchange chromatography medium, the cation exchange medium is Fastline SP; the phosphate buffer with pH 7.0 is passed through the bed from bottom to top, and the expanded bed is balanced. When the bed expands stably, the expansion rate is 2.0;

[0037] 3) Pass through the solution in step 1) for adsorption, then pass through the buffer used in step 2) to wash until no protein is washed out, then elute with pH 7.0 phosphate buffer containing 0.5 M NaCl, collect the eluted Liquid, its purity is 90%, and the recovery rate of lactoferrin is 83%;

[0038] 4) Wash the medium in place with 0.2 M NaOH and finally re-equilibrate the medium with pH 7.0 phosphate buffer.

[0039] In step 3), when feeding the solution in step 1) for adsorption, keep the operating flow rate the sam...

Embodiment 2

[0042] 1) Add water to dissolve sweet whey powder to make 1 L of whey solution with a mass percentage concentration of 20g / L, and adjust the pH to 8.0 with NaOH;

[0043] 2) Fill the expanded bed with cation exchange chromatography medium, the cation exchange medium is Fastline SP; use pH 8.0 phosphate buffer to pass through the bed from bottom to top, balance the expanded bed, and when the bed expands stably, the expansion rate is 2.6;

[0044] 3) Pass through the solution in step 1) for adsorption, then pass through the buffer used in step 2) to wash until no protein is washed out, then elute with phosphate buffer solution containing 1 M NaCl at pH 8.0, and collect the eluted Liquid, its purity is 93%, and lactoferrin recovery rate is 47%;

[0045] 4) Wash the medium in place with 0.2 M NaOH and finally re-equilibrate the medium with pH 7.0 phosphate buffer.

[0046] In step 3), when feeding the solution in step 1) for adsorption, keep the operating flow rate the same as the ...

Embodiment 3

[0048] 1) Add water to dissolve sweet whey powder to make 1 L of whey solution with a mass percentage concentration of 80g / L, and adjust the pH to 7.5 with NaOH;

[0049] 2) Fill the expanded bed with cation exchange chromatography medium, the cation exchange medium is Streamline SP; use pH 7.5 phosphate buffer to pass through the bed from bottom to top, balance the expanded bed, and when the bed expands stably, the expansion ratio is 1.8;

[0050] 3) Pass through the solution in step 1) for adsorption, then pass through the buffer used in step 2) to wash until no protein is washed out, then elute with pH 10.0 carbonate buffer containing 0.5 M NaCl, collect the washed Deliquification, its purity is 88%, and the recovery rate of lactoferrin is 86%;

[0051] 4) Wash the medium in place with 0.2 M NaOH and finally re-equilibrate the medium with pH 7.0 phosphate buffer.

[0052] In step 3), when feeding the solution in step 1) for adsorption, keep the operating flow rate the same...

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Abstract

The invention provides a method for purifying lactoferrin from milk serum by using an expanded bed adsorption technology. The method comprises the following steps of: (1) adding water into sweet whey powder for dissolving into a whey solution or adjusting the pH of commercially available liquid whey by using NaOH; (2) filling a cation exchanging chromatography medium into an expanded bed, making a phosphate buffer solution pass through a bed layer from bottom to top, and balancing the expanded bed; (3) introducing the solution obtained in the step (1) for adsorbing, eluting by using a buffer solution containing NaCl, and collecting an eluent; and (4) cleaning by using NaOH, and regenerating a medium by using a buffer solution. In the method, expanded bed adsorption is taken as a core separating method, the purity of lactoferrin obtained by one-step separation is up to 88-93 percent, the recovery ratio is 47-86 percent, a process is easy, convenient and efficient, the equipment investment is small, and the method is suitable for industrial production.

Description

technical field [0001] The invention relates to a method for purifying lactoferrin from whey by expanding bed adsorption technology. Background technique [0002] After the casein is separated from milk to make cheese, the remaining solution is whey. As a by-product, whey liquid has high water content and is inconvenient to transport. It is often made into whey powder by spray drying and other methods. The main ingredients in whey powder are lactose and whey protein. [0003] Whey proteins mainly include β-lactoglobulin (β-Lg), α-lactalbumin (α-La), bovine serum albumin (BSA), immunoglobulin (IgG), and lactoferrin (Lf), Lactoperoxidase, growth factors and many biologically active factors and enzymes all have certain biological activities, especially lactoferrin has attracted much attention because of its unique functions. [0004] The isoelectric point of lactoferrin is 8.0, the molecular weight is 80kDa, and the content in whey is about 0.5 g / L. Lactoferrin has many bio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/79C07K1/18
Inventor 姚善泾杜巧燕林东强
Owner ZHEJIANG UNIV
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