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Construction method of anti-naphthalene monoclonal antibody hybridoma cell line

A hybridoma cell line and monoclonal antibody technology, which is applied in chemical instruments and methods, biochemical equipment and methods, instruments, etc., can solve the problems of long time-consuming, high requirements for experimental equipment, cumbersome processing procedures, etc., and achieve specificity Strong, high potency effect

Active Publication Date: 2021-08-10
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 anti-naphthalene monoclonal antibody hybridoma cell line
  • Construction method of anti-naphthalene monoclonal antibody hybridoma cell line
  • Construction method of anti-naphthalene monoclonal antibody hybridoma cell line

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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 cou...

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Abstract

The invention discloses a method for constructing an anti-naphthalene monoclonal antibody hybridoma cell line; the method includes artificial antigen synthesis, mouse immunization, hybridoma cell screening and strain establishment, monoclonal antibody preparation, purification and potency Determination. The antigens include immunization antigens and detection antigens, and the screening and establishment of hybridoma cells include the collection of SP2 / 0 cells, the collection of splenocytes, cell fusion, and the detection of fusion plates. The preparation method of the present invention is simple, and the immune effect is good; the hybridoma cell line stably secreting anti-naphthalene monoclonal antibody can be obtained, and it is a more stringent method for subsequent establishment of the immune detection method for the interference of salinity and various ions in the water environment. The screening provides an experimental basis and a research basis for the rapid immunological 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 thing. 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 Patents(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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