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Universal phthalic acid esters monoclonal antibody hybridoma cell strain and application thereof

A technology of hybridoma cell line and phthalic acid, applied in the direction of anti-animal/human immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problems of cumbersome processing, expensive instruments, and complicated operations

Active Publication Date: 2014-08-27
无锡迪腾敏生物科技有限公司
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  • Abstract
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AI Technical Summary

Problems solved by technology

Among them, the instrument detection method is accurate, but the instrument is expensive, the operation is complicated, and the sample pretreatment is cumbersome.

Method used

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  • Universal phthalic acid esters monoclonal antibody hybridoma cell strain and application thereof

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Embodiment 1

[0036] Example 1 Preparation of Phthalate General Monoclonal Antibody

[0037] 1. Synthesis of hapten

[0038] 1.1 Esterification reaction: Weigh 0.12mol of 4-nitrophthalic acid into a 100mL three-necked flask, add 0.1mol of n-butanol, add 6mL of benzene to aid dissolution, stir magnetically, slowly add 1.6mL of concentrated sulfuric acid, and heat up to 120°C, react for 12h, evaporate the organic solvent under reduced pressure, add the oily residue into cold water and wash with 10% Na 2 CO 3 The solution was washed until the lower aqueous layer was colorless, and a dark red oily crude product was obtained in the upper layer, which was recrystallized with absolute ethanol to obtain 4-nitrophthalic acid dibutyl ester.

[0039] 1.2 Reduction reaction: Weigh 7 mmol of reduced iron powder into a 50 mL round bottom flask, add 2 mL of water and 1 mmol of ammonium chloride, and heat in a water bath for 15 min. Another 1.4 mmol of 4-nitrophthalate dibutyl was dissolved in 5 mL of m...

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Abstract

The invention discloses a universal phthalic acid esters monoclonal antibody hybridoma cell strain and application thereof, and belongs to the technical field of food safety immunological detection. Hapten is prepared and is coupled with albumen based on the glutaraldehyde method to obtain phthalic acid dibutyl ester complete antigen, and the phthalic acid dibutyl ester complete antigen and freund's adjuvant are evenly mixed to be injected to immune BALB / c mice in a subcutaneous injection mode; envelope antigen is formed in a synthesis mode based on the diazotization method and is used for screening mouse serum and cell supernatant. The splenocyte of the immune mice is fused with myeloma cells of the mice based on a PEG method, and indirect ELISA, indirect competition ELISA screening and three times of subcloning are carried out to obtain selective group hybridoma cell strain monoclonal cell strain C. The monoclonal cell strain C has certain recognition capability on DEHP and DINP, and the requirement for phthalic acid ester plasticizer immunodetection products in current market can be met.

Description

technical field [0001] The invention relates to a hybridoma cell line of a phthalate-type general-purpose monoclonal antibody and its application, relates to a preparation method of a phthalate-type plasticizer general-purpose monoclonal antibody, and belongs to the technical field of food safety immunology detection . Background technique [0002] A plasticizer is an additive that increases the softness of a material or liquefies a material. The most common plasticizers are Phthalate Esters (PAEs), which are esterified derivatives of phthalic acid. [0003] Phthalate plasticizers are colorless liquids with a little aromatic smell or no smell, medium viscosity, high stability, low volatility, low cost, low water solubility, but easily soluble in most organic solvents. Phthalates are widely used in daily life and industry, with di(2-ethylhexyl) phthalate (DEHP) as the largest, accounting for three-quarters of plasticizer production, followed by ortho Dibutyl phthalate (DBP...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/18G01N33/577C12R1/91
Inventor 匡华胥传来郭玲玲徐丽广马伟刘丽强宋珊珊吴晓玲
Owner 无锡迪腾敏生物科技有限公司
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