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Hybridoma cell strain No. 15, and pyrethroid-resistant mass selection monoclonal antibody produced by hybridoma cell strain No. 15

A hybridoma cell line and pyrethroid technology, which is applied in the field of immunochemistry, can solve the problems of high cost, complicated operators, and unsuitable for daily rapid detection of pyrethroid residues.

Active Publication Date: 2014-01-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods (except TLC) require expensive instruments, professional operators and complicated sample cleanup, and are not suitable for daily rapid detection of pyrethroid residues in agricultural products

Method used

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  • Hybridoma cell strain No. 15, and pyrethroid-resistant mass selection monoclonal antibody produced by hybridoma cell strain No. 15
  • Hybridoma cell strain No. 15, and pyrethroid-resistant mass selection monoclonal antibody produced by hybridoma cell strain No. 15

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Preparation of hybridoma cell line No. 15

[0019] 1. Animal immunity and serum screening

[0020] The conjugate of hapten 1 (H1) and hemocyanin (KLH) (H1-KLH) was selected as the immunogen, and the fast immune adjuvant and the immunogen were simply mixed according to a certain ratio, and BALB / c was immunized by intramuscular injection. For mice, the interval between immunizations was 2 weeks, and 7-10 days after the fourth immunization, the titer and inhibition of serum were measured by indirect competitive enzyme-linked immunosorbent assay, and mice with high titer and good inhibition were selected for fusion.

[0021] 2. Cell Fusion

[0022] The first step is to culture and collect tumor cells. 7-10 days before fusion, SP2 / 0 tumor cells were incubated with RPMI-1640 medium containing 10% FBS (fetal bovine serum) in 5% CO 2 in the incubator. The number of SP2 / 0 tumor cells is required to reach 1-4*10 before fusion 7 , while ensuring that the SP2 / 0 tumor...

Embodiment 2

[0027] Example 2 Preparation, purification, subtype and identification of anti-pyrethroid monoclonal antibody secreted by hybridoma cell line No. 15

[0028] 1. Preparation of monoclonal antibodies

[0029] Select healthy BALB / c mice and inject sterile paraffin oil at a volume of 0.6 mL / mouse. After 10 days, each mouse was intraperitoneally injected with 1*10 6 hybridoma cells. After the 6th day, observe the state of the mouse every day. If the abdomen of the mouse is obviously enlarged, feels tight when touched by hands, and is unwilling to move, collect ascites; then centrifuge (6000 rpm, 12 min) to remove red blood cells and other impurities, aliquoted and stored at -20°C.

[0030] 2. Monoclonal antibody purification

[0031]Ascitic fluid was purified by octanoic acid-ammonium sulfate precipitation. Under acidic conditions, n-octanoic acid can precipitate other miscellaneous proteins in ascites except IgG immunoglobulin, then centrifuge, and discard the precipitate; th...

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Abstract

The invention discloses hybridoma cell strain No. 15, and pyrethroid-resistant mass selection monoclonal antibody produced by the hybridoma cell strain No. 15, and belongs to the field of immunochemical technology.<{EN3}>The monoclonal antibody secreted by the cell strain is detected by indirect competition euzymelinked immunosorbent assay, and is capable of realizing cross reaction with monoclonal antibodies of 10 kinds of pyrethroids including fenpropathrin, cypermethrin, beta-cypermethrin, cyfluthrin, lambda-cyhalothrin, fenvalerate, deltamethrin, fluvalinate, cyphenothrin and alphamethrin; IC50 ranges from 3.2 to 50ng / ml; the pyrethroid-resistant mass selection monoclonal antibody produced by the hybridoma cell strain No. 15 can be used for development of colloidal gold immunochromatography test strips and immunosensors, is capable of providing raw materials for immunodetection of pyrethroid residue in agricultural products, and possesses practical application value. The monoclonal antibody secreted by the cell strain is detected by indirect competition euzymelinked immunosorbent assay, and is capable of realizing cross reaction with monoclonal antibodies of 10 kinds of pyrethroids including fenpropathrin, cypermethrin, beta-cypermethrin, cyfluthrin, lambda-cyhalothrin, fenvalerate, deltamethrin, fluvalinate, cyphenothrin and alphamethrin; IC50 ranges from 3.2 to 50ng / ml; the pyrethroid-resistant mass selection monoclonal antibody produced by the hybridoma cell strain No. 15 can be used for development of colloidal gold immunochromatography test strips and immunosensors, is capable of providing raw materials for immunodetection of pyrethroid residue in agricultural products, and possesses practical application value.

Description

technical field [0001] The invention relates to a mouse-derived hybridoma cell line No. 15 and a group-selective monoclonal antibody capable of simultaneously recognizing 10 kinds of pyrethroids, which can be used in the development of colloidal gold immunochromatographic test strips and immunosensors. It belongs to the technical field of immunochemistry. Background technique [0002] According to statistics, there are currently more than 70 kinds of pyrethroid insecticides in the world, accounting for 20% of the world's insecticide market; their insecticidal activity has increased by ten times or even a hundred times than before; The toxicity of insecticides to mammals and birds is very low. Therefore, the application of pyrethroid insecticides has been developed unprecedentedly, and agricultural production has also obtained huge economic benefits. However, it also brings the problem of pyrethroid residues. Although pyrethroid pesticides are environmentally friendly pesti...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/577
Inventor 匡华陈秀金徐利广胥传来宋姗姗任国艳张勋马伟
Owner JIANGNAN UNIV
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