Construction method of naphthalene resisting monoclonal antibody hybridoma cell strain

A hybridoma cell line, monoclonal antibody technology, applied in chemical instruments and methods, biochemical equipment and methods, instruments, etc., can solve the problems of high requirements for experimental equipment, long time consumption, low detection limit, etc., and achieve titer. High, specific effect

Active Publication Date: 2019-05-03
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these conventional detection methods can complete the qualitative or quantitative detection of PAHs, and can reach a lower detection limit, they have certain disadvantages, such as high requirements for experimental equipment, difficulty in on-site implementation, cumbersome sample pretreatment procedures, and costly long time

Method used

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  • Construction method of naphthalene resisting monoclonal antibody hybridoma cell strain
  • Construction method of naphthalene resisting monoclonal antibody hybridoma cell strain
  • Construction method of naphthalene resisting monoclonal antibody hybridoma cell strain

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Embodiment 1

[0033] This embodiment relates to a method for constructing an anti-naphthalene monoclonal antibody hybridoma cell line, said method comprising the steps of:

[0034] The first step, artificial antigen preparation

[0035] 2-Nabutyric acid was dissolved in DMSO to 50 mg / ml, KLH and OVA were dissolved in coupling buffer to 10 mg / ml, and EDC was dissolved in sterile water to 10 mg / ml.

[0036] Add 1.00mg (20ul) of 2-naphthoic acid to 250ul pH7.5 PBS coupling buffer, react at room temperature for 2h and place in a refrigerator at 4°C overnight, then add 100ul KLH (10mg / ml) and add 400ul EDC (10mg / ml), mix and rotate at room temperature for 2 hours, add 400ul PBS, and dialyze with 500ml PBS at 4°C for 72 hours in a dialysis bag. 2-Nabutyric acid-OVA was synthesized by the same method, and the coupling ratio of the artificial antigen was measured by spectrophotometer. As shown in Table 1.

[0037] Table 1 Conjugation ratio of artificial antigen

[0038] Couplet

co...

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Abstract

The invention discloses a construction method of a naphthalene resisting monoclonal antibody hybridoma cell strain. The method comprises the steps of synthetizing artificial antigens, performing mouseimmunization, screening hybridoma cells and establishing strains, performing preparation and purification of a monoclonal antibody, and performing potency determination, wherein the antigens comprisean immunization antigen and a detection antigen; and the step of screening hybridoma cells and establishing strains comprises the substeps of collecting SP2/0 cells, collecting spleen cells, combining cells, detecting a combining plate and the like. The preparation method disclosed by the invention is simple, and the immune effects are good; the hybridoma cell strain which can stably secrete naphthalene resisting monoclonal antibodies can be obtained, experiment foundation can be provided for subsequent stricter screening in accordance with factors of salinity, various ions and the like in water environment interfering establishment of an immunodetection method, and research foundation can be provided for research relevant to immunology fast detection of naphthalene in the water environment.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a method for constructing an anti-naphthalene monoclonal antibody hybridoma cell line. Background technique [0002] Naphthalene belongs to polycyclic aromatic hydrocarbons (PAHs), and is a raw material for the production of synthetic resins, plasticizers, rubber antioxidants, surfactants, synthetic fibers, dyes, paints, etc. The widely used naphthalene has become a typical environmental pollution things. In recent years, PAHs have also been detected to varying degrees in rivers and oceans. After people ingest PAHs in different ways, PAHs in the human body will directly or indirectly affect human health. A considerable portion of PAHs can cause diseases such as lung cancer, bladder cancer, and gastric cancer. At the same time, studies have shown that PAHs can cause damage to the human immune system, nervous system, and hematopoietic system. [0003] Among the 16...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/577
Inventor 周勇程金平胡雪马煜宁韩德明陈筱佳张旭峰
Owner SHANGHAI JIAO TONG UNIV
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