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Preparation method of mice antiserum resisting toad skin transgelin-2 recombinant protein

A technology of recombinant protein and gelatin, applied in the field of biomedicine, can solve the problems of less products, shortcomings of antiserum, and safety issues

Inactive Publication Date: 2013-11-27
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] To sum up, the conventional technology for the preparation of animal antiserum is relatively mature, but there are few products on the market at present, mainly due to safety issues, changes in antibody titers, and shortcomings of antiserum.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Mix toad transgelatin-2 recombinant protein and Freund's complete adjuvant at a ratio of 1:1 to form A agent; then toad transgelatin-2 recombinant protein and Freund's incomplete adjuvant at a ratio of 1:1 The ratio of the mixture was mixed into dose B, and then the mice were subcutaneously injected with 45ug of dose A, fed for 12 days, and then subcutaneously injected with dose B of 55ug, and then injected with dose B every 12 days, and repeated injections of dose B for 3 Then, blood was collected directly from the heart of the small white mouse, and the obtained blood was left to stand at 2°C for 15 hours. After the blood was fully coagulated, it was centrifuged at 2000 rpm for 35 minutes, and the supernatant was collected and stored in a -70°C ultra-low temperature refrigerator to obtain mouse antiserum.

Embodiment 2

[0024] Mix toad transgelatin-2 recombinant protein and Freund's complete adjuvant at a ratio of 1:1 to form A agent; then toad transgelatin-2 recombinant protein and Freund's incomplete adjuvant at a ratio of 1:1 The ratio of the mixture was mixed into dose B, and then the mice were subcutaneously injected with 100ug of dose A, fed for 14 days, then subcutaneously injected with dose B of 100ug, and then injected with dose B once every 14 days, and repeated injections of dose B for 5 Then, blood was collected directly from the heart of the small white mouse, and the obtained blood was left to stand at 6°C for 10 hours. After the blood was fully coagulated, it was centrifuged at 4000 rpm for 25 minutes, and the supernatant was collected and stored in a -70°C ultra-low temperature refrigerator to obtain mouse antiserum.

Embodiment 3

[0026] Mix toad transgelatin-2 recombinant protein and Freund's complete adjuvant at a ratio of 1:1 to form A agent; then toad transgelatin-2 recombinant protein and Freund's incomplete adjuvant at a ratio of 1:1 The ratio of the mixture was mixed into dose B, and then the mice were subcutaneously injected with 100ug of dose A, fed for 16 days, then subcutaneously injected with dose B of 45ug, and then injected with dose B every 12 days, and repeated injections of dose B for 4 Once, blood was collected directly from the heart of the small white mouse, and the obtained blood was left to stand at 3°C ​​for 11 hours. After the blood was fully coagulated, it was centrifuged at 3000 rpm for 30 minutes, and the supernatant was collected and stored in a -70°C ultra-low temperature refrigerator to obtain mouse antiserum.

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Abstract

The invention discloses a preparation method of a mice antiserum resisting a toad skin transgelin-2 recombinant protein and belongs to the technical field of biological medicine. The preparation method is characterized by comprising the following steps of 1, preparing a toad skin transgelin-2 recombinant protein and a Freund's complete adjuvant into an agent A, and preparing the toad skin transgelin-2 recombinant protein and a Freund's incomplete adjuvant into an agent B, 2, carrying out mice hypodermic multipoint injection of the agent A, carrying out normal feeding for 12-16 days, carrying out hypodermic injection of the agent B, and then carrying out injection of the agent B every 12-16 days, wherein the agent B injection process is totally carried out 3-5 times, and 3, collecting heart blood of the treated mice and carrying out centrifugation to obtain a supernatant which is the mice antiserum. The preparation method can be used for handling and forecasting specific events of transgelin-2 antitumor research and has effects of prompting and marking transgelin-2. The mice antiserum can be obtained simply, has a low cost and has obvious effects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a method for preparing mouse antiserum against toad transgelatin-2 recombinant protein. Background technique [0002] Antiserum (antiserum) is a serum containing polyclonal antibodies. Passive immunity can be delivered by injection of antiserum to treat many diseases. [0003] Two centuries ago, people injected the soluble toxin isolated from the diphtheria bacillus culture supernatant into horses, and the antiserum obtained could treat diphtheria. This is the first example of antibody treatment. After more than two hundred years of development, people have been able to obtain specific antibodies against toxins, viruses, bacteria and even cancer cells through various immune methods. In our country, the research and utilization of antiserum is relatively early. At the beginning of the last century, the Republic of China at that time planned to build the first serum institut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K16/06
Inventor 杨仙玉
Owner ZHEJIANG FORESTRY UNIVERSITY
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