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A hybridoma cell line of phthalates universal monoclonal antibody and its application

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: 2016-08-24
无锡迪腾敏生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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  • A hybridoma cell line of phthalates universal monoclonal antibody and its application

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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 relates to a hybridoma cell line of general monoclonal antibody of phthalates and its application, belonging to the technical field of food safety immunology detection. The present invention prepares hapten, couples hapten to protein by glutaraldehyde method, obtains dibutyl phthalate complete antigen, mixes with Freund's adjuvant evenly, and immunizes BALB / c mice by subcutaneous injection; The coated antigen was synthesized by nitrogenation method for screening of mouse serum and cell supernatant. Splenocytes from immunized mice were fused with mouse myeloma cells by PEG, screened by indirect ELISA and indirect competition ELISA and subcloned three times to obtain a group-selective hybridoma cell line monoclonal cell line C. The monoclonal cell line C provided by the present invention also has a certain ability to recognize DEHP and DINP, and can meet the current market demand for phthalate plasticizer immunoassay products.

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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Patent Type & Authority Patents(China)
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