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Method for measuring drug-loading rate of bacteria nano magnetosomes

A nano-magnetic and drug-loading technology, applied in color/spectral characteristic measurement, fluorescence/phosphorescence, material excitation analysis, etc., can solve problems such as drug decomposition and accurate determination of drug loading

Active Publication Date: 2008-05-07
南京英睿杰生化科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, for the bacterial nano-magnetosome system, these strong acids and strong polar solvents can easily change and decompose the properties of the loaded drug while dissolving and destroying the bacterial nano-magnetosome membrane and magnetic core, thereby affecting its drug loading. Accurate determination of quantity

Method used

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  • Method for measuring drug-loading rate of bacteria nano magnetosomes
  • Method for measuring drug-loading rate of bacteria nano magnetosomes

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1: The amount of anticancer drug doxorubicin (ADM) loaded on bacterial nano-magnetosomes (BMP) was determined by ultraviolet spectroscopy.

[0023] (1) Prepare a standard solution of doxorubicin: accurately weigh 0.02 g of doxorubicin, dissolve it in pure water, and prepare a standard solution in a 25 ml volumetric flask. Pipette different volumes of the above-mentioned doxorubicin standard solution in a 25ml volumetric flask, add water to dilute to the mark, and accurately prepare a series of doxorubicin standard solutions with known concentrations.

[0024] (2) Establish the standard working curve of the contained drug: use pure water as a reference, measure the ultraviolet scanning spectrum of the above-mentioned doxorubicin standard solution, and obtain its ultraviolet spectrum as shown in Figure 1. According to Figure 1, it can be determined that doxorubicin has the maximum absorption wavelength λ in pure aqueous solution max 480nm. Measure the absorbance...

Embodiment 2

[0030] Example 2: Detection of anti-CEA content of monoclonal antibody carried by bacterial nano-magnetosomes by fluorescence spectroscopy.

[0031] (1) Prepare anti-CEA standard solution: add phosphate buffer solution to prepare a series of anti-CEA standard solutions with known accurate concentrations.

[0032] (2) Establishing the standard working curve of anti-CEA: measure the fluorescence emission spectrum of the above-mentioned anti-CEA standard solution, and obtain the fluorescence emission spectrum as shown in FIG. 3 . Determine its maximum emission wavelength λ max 336nm. Measure the fluorescence emission intensity values ​​at 336 nm of the above-mentioned series of anti-CEA solutions with known accurate concentrations, and make the standard working curve as shown in FIG. 4 . Determine its regression equation as I=8.047*10 3* c. W is the absorbance value, and C is the concentration of the anti-CEA standard solution, that is, the absorbance value of the solution is...

Embodiment 3

[0037] Embodiment 3: Precision experiment

[0038] Taking fluorescence spectroscopy as an example, the same batch of anti-CEA solution and glutaraldehyde-modified bacterial nano-magnetosomes were subjected to parallel experiments under the same operating conditions as described above, and the bacterial nano-magnetic The amount of linked anti-CEA in the body, the results are shown in Table 1 below. This result shows that the detection method of the present invention can meet the ideal precision requirement.

[0039] Table 1 Results of precision experiments

[0040] Experiment number

[0041] Table 1 shows that the same batch of raw materials has been tested six times in parallel, and the error of drug loading measured is within the allowable range.

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Abstract

The invention relates to a way of determining the drug loading of the bacteria nanometer magnetosome, which uses an ultraviolet spectrum or a fluorescence spectrum to determine the biggest ultraviolet absorbing wavelength or biggest emission wavelength Gamma max of the tested drug. The ultraviolet absorbance values or the fluorescence transmission intensity values of the drug solution with different concentrations are tested at the Gamma max to draw the standard working curves. The drug with known concentration mixed-react with the bacteria nanometer magnetosome, and the magnetosome after reaction is separated to acquire the supernatant; and the drug solution with the same concentration and quantity with the former solution is prepared without reacting with the bacteria nanometer magnetosome to be as the comparison solution of the blank reaction; the ultraviolet absorbance value or the fluorescence transmission intensity value of the supernatant and the comparison solution of the blank reaction at the Gamma max are respectively determined, and the drug loading of the bacteria nanometer magnetosome is calculated by the standard working curves according to the difference of the two.

Description

technical field [0001] The invention relates to a method for measuring the drug-loaded amount of bacterial nano-magnetosomes, in particular to a method for indirectly measuring the drug-loaded amount of bacterial nano-magnetosomes by using ultraviolet spectrum or fluorescence spectrum. Background technique [0002] Magnetotactic bacteria (magnetotactic bacterium) is a kind of peculiar microorganism in nature that was accidentally discovered by the American biologist Blakemore in 1975. It can move along the magnetic field lines, and its cells contain magnetic field-sensitive Bacterial nano-magnetosomes (magnetosomes), which play a guiding role in the cell, can move with the help of their own flagella. The size of bacterial nanomagnetosomes is about 35-120nm, within the size range of permanent single magnetic domain crystals, and the main component is Fe 3 o 4 or Fe 3 S 4 , the outside is coated with phospholipid bimolecular lipid membrane of chimeric protein, after separa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33G01N21/64
Inventor 郭琳
Owner 南京英睿杰生化科技有限公司
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