A gold standard test strip or card for rapid detection of lead ions

A technology of gold standard test paper and lead ions, applied to gold standard test paper strip/card and its preparation method and application field

Inactive Publication Date: 2015-09-16
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the relevant literature and patents on the development of gold standard test strips / cards for rapid detection of lead ions have not yet been reported.

Method used

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  • A gold standard test strip or card for rapid detection of lead ions
  • A gold standard test strip or card for rapid detection of lead ions
  • A gold standard test strip or card for rapid detection of lead ions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Preparation of lead ion artificial immune antigen

[0036] Preparation of artificial immune antigen Pb using isothiocyanate method 2+ -ITCBE-OVA. Weigh 20 mg of ovalbumin OVA and dissolve it in 1 mL of pH 9.0 HEPES buffer (10 mM / L) to form a 20 mg / mL carrier protein solution. Weigh 10 mg of ITCBE and dissolve it in 1 mL of dimethyl sulfoxide (DMSO) to form a metal chelating agent solution; add 0.5 mL of metal chelating agent solution to the OVA carrier protein solution dropwise, and after mixing, add 100ul tri-n-butylamine ( 1.5M), react on a shaker at room temperature for 24 hours to prepare an OVA-ITCBE solution. Take 11.25mg of lead nitrate (Pb(NO 3 ) 2 ) Dissolve in 100ul distilled water to form uniform Pb 2+ Solution; Pb 2+ Add the solution to the OVA-ITCBE solution, adjust the pH to 7.4, incubate for 4 hours on a shaker at room temperature, and then transfer it into a dialysis bag for dialysis for 10 days to form Pb 2+ -ITCBE-OVA, collected and aliquoted,...

Embodiment 2

[0038] Example 2: Identification of lead ion artificial immune antigen

[0039] Determination of Pb by the quinolinic acid method 2+ -OVA concentration of carrier protein in ITCBE-OVA. With OVA as the standard protein, the concentration gradients of 0μg / mL, 10μg / mL, 20μg / mL, 40μg / mL, 60μg / mL, 80μg / mL were prepared, and the concentration detection standard curve was constructed by the bicinchoninic acid method. The linear equation of the OVA concentration standard curve is: y=0.0022x+0.008, R 2 =0.9987, where y is the absorbance of the sample at a wavelength of 562nm, and x is the protein concentration of the sample.

[0040] Determination of Pb by ICP-AES 2+ -Pb in ITCBE-OVA 2+ concentration. Add 100μg / mL Pb 2+ -Pb in ITCBE-OVA 2+ The standard stock solution is diluted with 2% nitric acid into concentration gradients of 0μg / mL, 0.1μg / mL, 0.2μg / mL, 0.3μg / mL, 0.4μg / mL, 0.5μg / mL, and the instrument software automatically draws the standard curve, and The linear regression equation Y...

Embodiment 3

[0041] Example 3: Preparation of anti-lead ion monoclonal antibody

[0042] Use Pb 2+ -ITCBE-OVA immunizes 5 female BALB / C mice for 6 weeks with a dose of 50μg / 0.2mL / mouse, injected subcutaneously on the back. Select cell fusion spare mice, indirect ELISA to detect Pb in antiserum 2+ Chelate pAb titer, block ELISA to detect Pb 2+ -ITCBE pAb vs. Pb 2+ -ITCBE's IC 50 , Select the highest potency, IC 50 The lowest mouse is super immune to cell fusion. The spleen was aseptically taken to prepare spleen cells using cell fusion technology, and fused with NS0 myeloma cells under the action of 50% PEG (pH 8.0); positive hybridoma cell lines were screened by indirect ELISA and blocking ELISA, and strong positive was selected , The wells with high inhibition rate and vigorous cell growth were cloned by limiting dilution for 3 times. The hybridoma cells were resuscitated after 15 days, 30 days and 60 days of cryopreservation, and the supernatant was taken after culture. The antibody titer ...

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Abstract

The invention discloses a gold-labeled test strip / card used for quick detection of lead ions. The gold-labeled test strip / card comprises a bottom layer support plate (1), a sample cushion (2), a gold-labeled antibody combination cushion (3), a cellulose membrane (4) and a water absorption cushion. The gold-labeled test strip / card can realize the quick detection for contamination residuals of the lead ions in environment, soil, water, food, medicine, and cosmetics.

Description

Technical field: [0001] The invention relates to a gold standard test strip / card for rapid detection of lead ions and a preparation method and application thereof, belonging to the technical field of immunology and sanitary inspection. Background technique: [0002] Lead (Pb) is a blue-gray heavy metal, one of the environmental poisons we know well, it is widely distributed in nature, and it is one of the non-ferrous metals with multiple uses in modern society. As early as the Xia and Shang periods, our ancestors used copper and lead to smelt bronze with high hardness and toughness, which was widely used in the production of weapons, tools, ceremonial vessels and food utensils. The lead content of bronzes can reach about 40%. With the development of industry and agriculture, the use of lead has become more and more extensive, so that it can be found in the air, soil, food, tableware, and toys. In my country, the output of lead and refined lead ranks third in the world, mainly us...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/544G01N33/532
CPCG01N33/532G01N33/544G01N33/558G01N33/577
Inventor 葛亚明王自良范国英王爱萍张海棠张慧辉
Owner HENAN INST OF SCI & TECH
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