Preparation method of bisphenol A-coated antigen with polylysine as carrier and application thereof

A technology of polylysine and antigen-coating, which is applied in the field of ELISA, can solve the problems of no bisphenol A artificial packaging, etc., and achieve the effect of facilitating coupling reaction, simple structure and wide sources

Active Publication Date: 2015-02-04
深圳市绿诗源生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no patents on the preparation of bisphenol A artificially coated antigens using polylysine as a carrier.

Method used

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  • Preparation method of bisphenol A-coated antigen with polylysine as carrier and application thereof
  • Preparation method of bisphenol A-coated antigen with polylysine as carrier and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Weigh 61 mg of bisphenolic acid (BVA) and dissolve it in 2 mL of DMF; then weigh 30.6 mg of NHS and 53.2 mg of EDC·HCL and dissolve them in 2 mL of (tetrahydrofuran) DMF, slowly add it dropwise to the BVA solution, and stir at room temperature for 24 hours in the dark. The supernatant was collected by centrifugation to obtain liquid A.

[0028] Weigh 126 mg of poly-L-lysine and dissolve it in 5 mL of pH=7 phosphate buffer solution to form B solution, take 4 mL of A solution and slowly add dropwise to B solution, stir at room temperature for 24 hours in the dark, centrifuge, and transfer the supernatant to The dialysis bag was dialyzed with a phosphate buffer solution of pH = 7.4 for 72 hours, and the dialysate was changed every 12 hours. Finally, the dialysate was freeze-dried to obtain the artificially coated antigen of bisphenol A, that is, PLL-BVA, and its structure was identified by ultraviolet spectroscopy. refer to figure 1 .

Embodiment 2

[0030] Weigh 20.7 mg of bisphenolic acid (BVA) and dissolve it in 1 mL of DMF; then weigh 6.6 mg of NHS and 14.2 mg of EDC·HCL and dissolve it in 1 mL (tetrahydrofuran) DMF, slowly add it dropwise to the BVA solution, and stir for 2 hours at room temperature in the dark , and centrifuged to get the supernatant to obtain liquid A.

[0031] Weigh 30.2 mg of poly-mixed lysine and dissolve it in 5 mL of phosphate buffer solution with pH=7 to form liquid B, take 3 mL of liquid A and slowly add it dropwise to liquid B, stir at room temperature for 2 hours in the dark, centrifuge, and transfer the supernatant To the dialysis bag, dialyze with pH = 7.4 phosphate buffer solution for 72 hours, change the dialysate every 12 hours, and finally freeze-dry the dialysate to obtain bisphenol A artificial coating antigen, namely PLL-BVA, and identify its structure by ultraviolet spectroscopy , see figure 1 .

Embodiment 3

[0033]Weigh 24 mg of bisphenolic acid (BVA) and dissolve it in 1 mL of DMF; then weigh 10 mg of NHS and 17.2 mg of EDC·HCL and dissolve them in 1 mL (tetrahydrofuran) DMF, slowly add it dropwise to the BVA solution, and stir at room temperature for 12 hours in the dark. The supernatant was collected by centrifugation to obtain liquid A.

[0034] Weigh 15 mg of poly-L-lysine and dissolve it in 5 mL of pH=7 phosphate buffer solution to form B solution, take 2 mL of A solution and slowly add dropwise to B solution, stir at room temperature in the dark for 12 hours, centrifuge, and transfer the supernatant to The dialysis bag was dialyzed with a phosphate buffer solution of pH = 7.4 for 72 hours, and the dialysate was changed every 12 hours. Finally, the dialysate was freeze-dried to obtain the artificially coated antigen of bisphenol A, that is, PLL-BVA, and its structure was identified by ultraviolet spectroscopy. refer to figure 1 .

[0035] Bovine serum albumin was selected ...

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Abstract

The present invention discloses a preparation method of bisphenol A-coated antigen with polylysine as a carrier and application thereof, is aimed at providing the preparation method of bisphenol A-coated antigen with polylysine as the carrier which is simple in the synthetic method and the application thereof, and finally establishes an enzyme linked immunosorbent assay kit. The kit has the advantages of simple pre-treatment, fast speed and accuracy and low cost, and can be used for on-site testing in large scale. The technical points include: taking polylysine as a carrier protein, preparing bisphenol A-coated antigen by coupling with diphenolic acid, and applying the bisphenol A-coated antigen to prepare the bisphenol A enzyme linked immunosorbent assay kit, wherein the bisphenol A enzyme linked immunosorbent assay kit is composed of a solid phase carrier of envelope antigen, an antibody working solution, a bisphenol A standard solution, an enzyme labeled secondary antibody solution, an antibody diluent, a wash concentrated solution, a substrate solution and a stop solution. The preparation method belongs to the technical field of enzyme linked immunoassay.

Description

technical field [0001] The invention relates to a method for preparing a bisphenol A-coated antigen with polylysine as a carrier and an application thereof, and belongs to the technical field of enzyme-linked immunity. Background technique [0002] Bisphenol A (Bisphenol A, BPA) is 2, 2-bis (4-hydroxyphenyl) propane, which imitates or interferes with endogenous estrogen in the body by binding to estrogen receptors or affecting cell signal transduction pathways, etc. Irritant to human skin, respiratory tract, digestive tract and cornea. When the bisphenol A in the body reaches a certain amount, it will destroy the liver cells and kidney cells, causing chronic poisoning, causing people to experience symptoms such as dizziness, headache, rash, nervousness and diarrhea in varying degrees. In recent years, bisphenol A, which has estrogen-like effects, has been found in barreled drinking water, baby bottles and various water bodies around the world (Science. 2007,317(5840):884-88...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/531
Inventor 赵肃清周剑青张磊张焜周丽华
Owner 深圳市绿诗源生物技术有限公司
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