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Hybridoma cell strain capable of excreting monoclonal antibody of human carboxylesterase 2 and monoclonal antibody of human carboxylesterase 2

A technology of hybridoma cell lines and monoclonal antibodies, applied in anti-enzyme immunoglobulin, biochemical equipment and methods, microorganisms, etc., can solve the problems of unexplored physiological functions and unclear natural substrates of carboxylesterases, etc.

Inactive Publication Date: 2008-09-17
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Although many scientific and technical personnel have been working hard to study human carboxylesterase 2 in recent years, and there are some related research reports, the natural substrates of carboxylesterase (including carboxylesterase 2) are still unclear, and their physiological effects are still to be determined. explore

Method used

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specific Embodiment approach 1

[0004] Specific embodiment 1: The hybridoma cell line secreting human carboxylesterase 2 monoclonal antibody in this embodiment is prepared by the following steps: (1) Preparation of immunogen: extracting human normal liver, crushing and centrifuging to prepare liver microsomal protein (2) Animal immunization: the emulsified liver microsomal protein was injected into the back and abdominal cavity of the mice, and the immunization was repeatedly boosted until the antibody titer was equal to or higher than 1:1000; (3) Preparation of trophoblasts: HAT cultured The liquid was injected into the abdominal cavity of normal mice. After kneading the abdomen of the mice for 1-3 min, the liquid in the abdominal cavity was withdrawn and centrifuged. The centrifuged precipitate was added with HAT medium into the well culture plate, and cultured at 37±0.5 °C; (4) Preparation of splenocytes. : The booster immunized mice were injected with the same dose of human liver microsomal protein as the...

specific Embodiment approach 2

[0006] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the mice are 4-12-week-old BALB / c (inbred) female mice. Others are the same as the first embodiment.

specific Embodiment approach 3

[0007] Embodiment 3: The difference between this embodiment and Embodiment 1 is that: in step (2), the first booster immunization injection is performed in the third week after the first immunization injection, and thereafter, the booster immunization injection is performed every two weeks. The dose of booster immunization injection is the same as that of first immunization injection; the first immunization injection is emulsified with Freund's complete adjuvant, and Freund's incomplete adjuvant is used for booster immunization instead of Freund's complete adjuvant. Other steps are the same as those in the first embodiment.

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Abstract

This invention relates to a monoclonal antibody of human carboxylesterase 2 and the cell line of hybridoma secreting thereof, providing a powerful tool for the futher study of human carboxylesterase 2. The preparation of cell line comprises: (1)preparation of antigens, (2)animal immune, (3)preparation of nourishing cells, (4)preparation of spleen cells, (5) pretreatment of myeloma cells, (6)mixing, (7) cell fusion, (8)culture of the fused cells, (9)clonizing of positive hybridoma cells. The preparation of antibodies comprises: (1)inoculating of hybridoma cells, (2)collecting the ascites containing the antibodies, (3)purifying the antibodies, (4)dialyzing.

Description

technical field [0001] The present invention relates to a hybridoma cell line and a monoclonal antibody. Background technique [0002] Although many scientists have been working hard to study human carboxylesterase 2 in recent years, and there are some related research reports, the natural substrates of carboxylesterases (including carboxylesterase 2) are still unclear, and their physiological roles are still pending. explore. SUMMARY OF THE INVENTION [0003] The purpose of the present invention is to prepare hybridoma cell lines that secrete human carboxylesterase 2 monoclonal antibody and human carboxylesterase 2 monoclonal antibody, which provides a powerful tool for further research on human carboxylesterase 2 . The hybridoma cell line secreting human carboxylesterase 2 monoclonal antibody was prepared by the following steps: (1) preparation of immunogen: extracting human normal liver, crushing and centrifuging to prepare liver microsomal protein; (2) animal immuniz...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/18C12P21/08C07K16/40
Inventor 朱大岭唐晓波武正华王志刚纪宏宇于磊
Owner HARBIN MEDICAL UNIVERSITY
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