Construction method of peak camel phage display nano antibody

A phage display and nanobody technology, applied in the field of biomedicine, can solve problems such as death, side effects of anemia, red blood cell agglutination, etc., and achieve the effect of small impact on the human body, promoting phagocytic ability, and avoiding the inhibition of phagocytosis.

Active Publication Date: 2022-07-08
杭州荣谷生物科技有限公司
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Problems solved by technology

However, further studies have found that human red blood cells also express a large amount of CD47 protein. Therefore, after ordinary

Method used

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  • Construction method of peak camel phage display nano antibody
  • Construction method of peak camel phage display nano antibody
  • Construction method of peak camel phage display nano antibody

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Experimental program
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Effect test

Embodiment

[0114] S1. Construction of antibody library;

[0115] S11. Select healthy Bactrian camels for immunization, and collect 5 mL of blood before the initial immunization and reserve as negative serum;

[0116] S12. Initial immunization:

[0117] CD47 antigen was selected, and complete Freund's adjuvant was mixed with the antigen 1:1. After emulsification, the camel was subcutaneously injected at multiple points, and the inoculation amount was 0.5 mg of antigen per camel; after inoculation, 5 ml of peripheral blood was collected to detect its immune response. ;

[0118] S13. The second immunization:

[0119] The CD47 antigen was mixed with complete Freund's adjuvant 1:1, and after emulsification, the camels were injected subcutaneously at multiple points, and the inoculation amount was 0.2 mg of the antigen per camel; after inoculation, 5 ml of peripheral blood was collected to detect the immune response;

[0120] S14. The third, fourth and fifth immunizations:

[0121] Repeat ...

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Abstract

The invention discloses a construction method of a peak camel source phage display nano antibody, which comprises the following steps: based on a phage display technology, preparing an antibody by utilizing peak camel immunization, preparing the antibody, loading the antibody to a phage display carrier, and screening the prepared antibody by utilizing the characteristic that a specific antibody can be obtained by screening by utilizing a phage display antibody library, so as to obtain the peak camel source phage display nano antibody. The hCD47nb nano antibody with good binding capacity aiming at CD47 is obtained, the prepared hCD47nb can effectively promote the phagocytic ability of macrophages to cancerous cells, inhibition of the CD47 to the phagocytic effect of the macrophages is avoided, meanwhile, the prepared hCD47nb nano antibody is basically not combined with human red blood cells, aggregation of the red blood cells cannot be caused, the influence on the human body is small, and the hCD47nb nano antibody has good application prospects. Therefore, the cable has excellent use performance and market prospect.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for constructing a camel-derived phage-displayed nanobody. Background technique [0002] In recent years, cancer has gradually become one of the malignant diseases that seriously endanger the health of the Chinese people, and the incidence rate has repeatedly hit new highs. However, among the conventional cancer treatment methods, surgical resection has the most direct and quickest curative effect, but there are risks during the operation. At the same time, it is only suitable for the early stage of cancer, and the operation of sensitive parts and vital organs is dangerous and the success rate is low; Although chemotherapy does not face the risk of surgery, it has the disadvantages of being time-consuming and having large side effects on the human body. In recent years, targeted therapy, which has gradually emerged, has gradually entered people's field of vision due ...

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Application Information

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IPC IPC(8): C07K16/28C07K16/06C12N15/63
CPCC07K16/2803C07K16/06C12N15/63C07K2317/569Y02A50/30
Inventor 钱文斌叶倩王世兵陈洁徐云飞
Owner 杭州荣谷生物科技有限公司
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