Method for reducing complement level and protein precipitation by premixing bovine serum raw materials

A pre-mixed, serum technology, applied in the field of serum, to achieve the effect of reducing complement activity, good correspondence, and sufficient effect and time

Active Publication Date: 2020-06-09
RONG YE LANZHOU BIOLOGIC TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present inventor was surprised to find that the above-mentioned problems can be effectively solved at the same time by fully premixing bovine serum raw materials at low temperature, which has never been found and implemented in the art

Method used

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  • Method for reducing complement level and protein precipitation by premixing bovine serum raw materials
  • Method for reducing complement level and protein precipitation by premixing bovine serum raw materials
  • Method for reducing complement level and protein precipitation by premixing bovine serum raw materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Thorough premixing process

[0036] Blood was collected from newly born newborn calves by sealing the artery with a six-pack bag. After the blood enters the six-pack bag, it is coagulated, centrifuged at 4000rpm for 30 minutes, and the supernatant is taken by siphon, which is the raw material of bovine serum. Each bag of bovine serum should be marked with the source, blood collection time and operator. After being transported to the production factory through the cold chain, the bovine serum in the blood collection tube (utility model patent has been applied for) is taken out for quality inspection, and the virus (specifically, BVDV, phage, etc.), antibody (specifically, foot-and-mouth disease antibody, IgG) etc. Screening for multiple metrics.

[0037]Qualified raw materials are screened and pre-mixed with serum raw materials of 100 newborn cows, and mixed thoroughly at low temperature (10-15°C, specifically, 12°C) for 2-3 hours (specifically, 2.5 hours) (...

Embodiment 2

[0039] After the raw serum is fully premixed, the determination of the complement activity of the filtered finished serum and the serum that is not fully premixed is compared.

[0040] 1. Prepare goat antiserum and dilute it to achieve the best sensitization state.

[0041] (1) Rabbit red blood cells (1×10 9 cells / mL) subcutaneously into adult goats, 3ml each time, once a week, for a total of 6 weeks.

[0042] (2) Collect blood from jugular vein of goats at intervals of four days after stopping injection, and separate serum.

[0043] (3) Heat at 56°C for 30 minutes to inactivate the endogenous complement in the antiserum.

[0044] (4) Use GVB2+ (gelatin veronal buffer, gelatin veronal buffer; pH=7.4) solution to make two-fold serial dilutions of the antiserum from 1:100 to 1:1600.

[0045] (5) Take 0.1 mL of the diluent and mix it with the rabbit erythrocyte suspension at a ratio of 1:1, and place the mixture in a water bath at 37°C for 30 minutes. Add 1.3mL of 10-fold dil...

Embodiment 3

[0063] After the raw serum was premixed for a period of time, protein precipitation occurred. These protein precipitates are blocked with a filter cloth before filtering, and then filtered.

[0064] see figure 1 , This figure reflects the protein precipitation situation, wherein, A picture is the protein precipitation situation after mixing; B picture is the protein precipitation situation after the mixing blocks the precipitation.

[0065] see figure 2 , which shows the filtered finished product after thawing. Among them, the picture on the left is the finished product that is fully pre-mixed at low temperature and filtered, and the picture on the right is the finished product that is filtered after rapid mixing.

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Abstract

The invention relates to the technical field of bovine serum, and improves the quality of bovine serum products by fully mixing bovine serum at low temperature in advance. A bovine serum raw materiallow-temperature sufficient premixing process is innovatively adopted, and the problem that bovine serum effective components are severely damaged due to the fact that a bovine serum complement systemneeds to be treated at 56 DEG C for 30 minutes during elimination is solved. When bovine serum of different blood types is fully mixed by using the bovine serum premixing process, protein is separatedout, and if the bovine serum is only mixed before final filtration, the reaction time is insufficient, and the protein separation phenomenon of the final product is common, the bovine serum preparedby adopting the process can effectively avoid the protein separation. The method also has an advantage that the raw materials are fully premixed and then subjected to preliminary screening of detection items so that the method has better correspondence with the detection items of the final product.

Description

technical field [0001] The invention relates to the technical field of serum, in particular to a method for fully premixing bovine serum raw materials at low temperature. The invention fully mixes the bovine serum at low temperature in advance to reduce the negative impact of the complement system in the serum; solves the problem of protein precipitation that is easy to occur in the final product of bovine serum; at the same time, the pre-mixing process can also pre-mix the raw materials before testing items The preliminary screening of this kind of raw material corresponds better to the testing items of the final product. Background technique [0002] Blood contains plasma and blood cells. Blood is serum after removing blood cells and fibrin. The main function of serum is to carry blood cells, transport substances needed to maintain human life activities and waste generated in the body, etc. Serum is equivalent to the intercellular space of connective tissue. Serum is an i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/38G01N1/40G01N1/42
CPCG01N1/28G01N1/38G01N1/4077G01N1/42
Inventor 陈亮任志斌李晔陈强王元
Owner RONG YE LANZHOU BIOLOGIC TECH
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