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Method for obtaining natural prealbumin from serum

A prealbumin and serum technology, applied in the field of methods and its preparation, can solve the problems of target protein denaturation, complex steps, slow speed, etc., and achieve the effect of optimized removal steps, simple and fast operation

Active Publication Date: 2017-11-03
SICHUAN XINCHENG BIOLOGICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is: in the prior art, the steps of removing the Alb protein in the PA protein are complicated, the speed is slow, or the problem of denaturation of the target protein is caused. The purpose is to provide a method for obtaining natural prealbumin from serum. The method, through the optimization of reagents and process steps, can effectively achieve the purpose of quickly removing the Alb protein in the PA protein while avoiding the denaturation of the target protein, and the method has low cost and remarkable effect

Method used

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  • Method for obtaining natural prealbumin from serum
  • Method for obtaining natural prealbumin from serum
  • Method for obtaining natural prealbumin from serum

Examples

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Embodiment 1

[0028] A method of obtaining natural prealbumin from serum comprising:

[0029] a. Centrifuge the waste human serum to remove fat, take 100ml;

[0030] b. Dilute hydrochloric acid to adjust the serum pH to 4.80;

[0031] c. Add 8ml n-octanoic acid, the Alb protein precipitates;

[0032] d. Centrifuge at 18000r / min at 4°C for 20min at high speed, and filter through a 0.65μm PP membrane to obtain the filtrate;

[0033] e. Adjust the pH value of the filtrate to 4.60, add 9% PEG4000, the PA protein aggregates to form a light blue precipitate, and obtain PA

[0034] f. PA protein is precipitated and reconstituted, then dialyzed to remove impurities and concentrated to make a finished product. The concentration factor is 10 times, and the dialysate contains 10% PEG20000.

[0035] The contents of PA protein, Alb protein and TP protein in the finished product were determined, and the detection results are shown in Table 1.

[0036] Table 1

[0037]

[0038] It can be seen from...

Embodiment 2

[0040] The difference between this embodiment and embodiment 1 is that the parameters are different in this embodiment, and the specific preparation method of this embodiment is as follows:

[0041] a. Centrifuge the waste human serum to remove fat, take 100ml;

[0042] b. Dilute hydrochloric acid to adjust the serum pH to 4.30;

[0043] c. Add 3ml n-octanoic acid, the Alb protein precipitates;

[0044] d. Centrifuge at 18000r / min at 4°C for 20min at high speed, and filter through a 0.65μm PP membrane to obtain the filtrate;

[0045] e. Adjust the pH value of the filtrate to 4.30, add 3% PEG4000, the PA protein aggregates to form a light blue precipitate, and obtain the PA protein precipitate;

[0046]f. PA protein was precipitated and reconstituted, then dialyzed to remove impurities and concentrated 2.5 times, and the dialysate contained 3% PEG20000.

Embodiment 3

[0048] The difference between this embodiment and embodiment 1 is that the parameters are different in this embodiment, and the specific preparation method of this embodiment is as follows:

[0049] a. Centrifuge the waste human serum to remove fat, take 100ml;

[0050] b. Dilute hydrochloric acid to adjust the serum pH to 4.60;

[0051] c. Add 5ml n-octanoic acid, the Alb protein precipitates;

[0052] d. Centrifuge at 18000r / min at 4°C for 20min at high speed, and filter through a 0.65μm PP membrane to obtain the filtrate;

[0053] e. Adjust the pH value of the filtrate to 4.40, add 5% PEG4000, the PA protein aggregates to form a light blue precipitate, and obtain the PA protein precipitate;

[0054] f. PA protein was precipitated and reconstituted, then dialyzed to remove impurities and concentrated 2.5 times. The dialysate contained 5% PEG20000.

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Abstract

The invention discloses a method for obtaining natural prealbumin from serum and solves a problem that complex manners and steps and low speed of removal of Alb protein from PA protein can cause denaturation of a target protein in the prior art. The method provided by the invention comprises the following steps: (1) after filtering serum to remove fat, adjusting a pH value to 4.60-4.90, adding an Alb specific binding substance to precipitate Alb, and collecting filtrate after centrifuging and filtering; (2) lowering a pH value of the filtrate to 4.40-4.70, adding a PA protein coagulant to obtain PA protein precipitates; (3) precipitating and re-dissolving the PA protein precipitates, and dialyzing to obtain a finished product. The method provided by the invention can effectively achieve a purpose of rapidly removing the Alb protein in the PA protein, and has advantages of low cost, significant effect and the like.

Description

technical field [0001] The invention relates to a kit and a preparation method thereof, in particular to a method for obtaining natural prealbumin from serum and a preparation method thereof. Background technique [0002] Recombinant PA has high purity, but it is different from natural PA in antigenic determinant and protein structure, which leads to different recognition of recombinant PA and natural PA by PA antibody, and recombinant PA is expensive, so high concentration of natural PA can be obtained from serum Albumin is one of the most effective ways. However, due to the similar isoelectric point and molecular weight of PA protein and Alb protein, it is difficult to distinguish the two proteins in the separation and purification process in the prior art. Protein assay experiments found that in the complex of Alb and PA, with the increase of Alb protein concentration The measured value of PA protein concentration decreased gradually with increasing gradually. Therefore...

Claims

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Application Information

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IPC IPC(8): C07K14/76C07K1/36C07K1/34C07K1/30
CPCC07K14/76
Inventor 徐建建林源谭韬魏建波
Owner SICHUAN XINCHENG BIOLOGICAL CO LTD
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