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Method for measuring content of IgG (immunoglobulin G) in pig plasma protein powder

A technology of porcine plasma protein powder and determination method, which is applied in the field of IgG content determination, can solve the problems of affecting the measurement result and incomplete IgG extraction, and achieves the effect of convenient operation, good extraction effect and good linear relationship

Inactive Publication Date: 2018-10-23
ZHANGZHOU AONONG ANIMAL HUSBANDRY TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for a powder sample such as porcine plasma protein powder, if the IgG content is measured using the method specified in the national standard, the sample may be attached to the blade of the homogenizer during the homogenization process, and the processing time is only 5 minutes. It will lead to incomplete extraction of IgG and affect the final measurement results

Method used

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  • Method for measuring content of IgG (immunoglobulin G) in pig plasma protein powder
  • Method for measuring content of IgG (immunoglobulin G) in pig plasma protein powder
  • Method for measuring content of IgG (immunoglobulin G) in pig plasma protein powder

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Effect test

Embodiment 1

[0027] The determination method of IgG content in pig plasma protein powder is:

[0028] Accurately weigh about 0.1000g of porcine plasma protein powder, and add it to 50mL of potassium phosphate buffer solution with pH=6.5 and 0.05mol / L (i.e., the ratio of solid to liquid is 1:500); Shake at a rotating speed for 30 minutes for constant temperature extraction, then centrifuge at 8000r / min for 10 minutes, and pass through a 0.22 μm filter membrane to obtain the test solution; take 20 μL of the test solution for high performance liquid chromatography detection.

[0029] Wherein, the high performance liquid chromatography conditions are as follows:

[0030] (1) Equipment: Agilent 1260, equipped with a UV detector, the detection wavelength is 280nm;

[0031] (2) Chromatographic column: Pharmacia HI-Trap Protein G column, 1mL;

[0032] (3) Mobile phase A: pH 6.5, 0.05mol / L potassium phosphate buffer (0.05mol / L potassium dihydrogen phosphate and 0.05mol / L dipotassium hydrogen phos...

Embodiment 2

[0043] The determination of IgG content in pig plasma protein powder is as follows:

[0044] 1. Solvents and equipment required for the test

[0045] 1.1HPLC: Aglient 1260, with UV detector;

[0046] 1.2 Chromatographic column: Pharmacia HI-Trap Protein G column, 1mL;

[0047] 1.3Millipore pure water machine;

[0048] 1.4 Analytical balance: accurate to 0.0001g;

[0049] 1.5 Erlenmeyer flask with stopper: 250mL;

[0050] 1.6 Pipette: 50mL pipette, which needs to be calibrated regularly;

[0051] 1.7 Constant temperature shaking incubator: Shanghai Jinghong HZP-150;

[0052] 1.8 high-speed centrifuge: the speed can be adjusted from 0 to 16000r / min;

[0053] 1.9 Mobile phase A: pH 6.5±0.1, 0.05mol / L potassium phosphate buffer (68.5% 0.05mol / L potassium dihydrogen phosphate and 31.5% 0.05mol / L dipotassium hydrogen phosphate are mixed evenly, and fine-tuned with a pH meter);

[0054] 1.10 Mobile phase B: pH 2.5±0.1, 0.05mol / L glycine hydrochloride buffer (250mL 0.2mol / L gly...

Embodiment 3

[0089] A method for assaying IgG content in pig plasma protein powder, comprising the following steps:

[0090] 1) Sampling: Weigh porcine plasma protein powder and add it to potassium phosphate buffer, add 2g of porcine plasma protein powder to every 1000mL potassium phosphate buffer to obtain the sample to be tested, wherein the concentration of potassium phosphate buffer is 0.05mol / L, the pH value is 6.5;

[0091] 2) Constant temperature extraction: place the sample to be tested in a constant temperature shaker at 25°C for 30 minutes at a speed of 150r / min, then centrifuge at 8000r / min for 10 minutes, and then filter the centrifuged sample to be tested with a 0.22μm filter membrane , to obtain the liquid to be tested;

[0092] 3) Get the liquid to be tested and carry out high-performance liquid chromatography detection. During the high-performance liquid chromatography detection process, mobile phase A is a phosphate buffer (pH value is 6.5), and mobile phase B is a glyci...

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Abstract

The invention relates to a method for measuring content of IgG (immunoglobulin G) in pig plasma protein powder. The method comprises the following steps of (1) sampling: weighing the pig plasma protein powder and adding into an extracting solution, so as to obtain the to-be-detected sample; (2) constant-temperature extracting: putting the to-be-detected sample into a constant-temperature rocking bed or an oscillation culture box, extracting, centrifuging, and filtering, so as to obtain the to-be-detected liquid; (3) detecting the to-be-detected liquid by high performance liquid chromatography.Compared with the prior art, the method has the advantages that the extracting effect is better, the accuracy of measuring result is high, the data stability is good, the operation is rapid, the linear relationship is good, and the method is suitable for being popularized and applied.

Description

technical field [0001] The invention belongs to the technical field of measuring IgG content, and relates to a method for measuring IgG content in pig plasma protein powder. Background technique [0002] my country is rich in pig blood resources, and the total amount of pig blood is about 10 per year. 8 kg. As a high-quality protein feed material, pig plasma protein powder retains the activity of functional immunoglobulin (Ig, a type of globulin with antibody activity or chemical structure similar to antibody) in the original plasma during its production process. These active substances can significantly reduce immune stimulation and effectively prevent post-weaning growth stagnation and diarrhea in piglets. [0003] Immunoglobulin (Ig) can be divided into IgG, IgA, IgM, IgD, IgE, etc. according to different antigenicity. IgG is the most important antibody in animal serum and plays the role of "main immunity" in humoral immunity. However, in the process of pig blood coll...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 王少青傅兴震吴有林陈淋敏杨伟春张军傅心锋周通杨再龙
Owner ZHANGZHOU AONONG ANIMAL HUSBANDRY TECH
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