Preserving fluid and preparation method thereof

A technology of preservation solution and carrageenan, which is applied in the field of chemiluminescence immunoassay, can solve the problems of affecting test results, unfavorable stable storage of markers on particles, and affecting antibody immune response

Inactive Publication Date: 2016-02-03
SINOCARE
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, with the rapid development of chemiluminescence immunoassay technology, many shortcomings have also been exposed. One of them is the preservation of magnetic particles and magnetic nanoparticles. If the viscosity of the preservation solution of magnetic particles or magnetic nanoparticles in the test strip is too low, First of all, magnetic particles or magnetic nanoparticles are prone to agglomeration and sinking to the bottom, which is not conducive to the stable storage of markers on the particles. Secondly, during transportation or use, they are likely to splash and stick to the wall or top of the container, thus making the detection process difficult. The number of particles that play a role in the medium decreases, which affects the test results; on the contrary, if the viscosity is too high, the problem of storage may be solved, but because the viscosity is too high, it is not conducive to the uniform dispersion of the mixed liquid, which will affect the antibody immune response

Method used

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  • Preserving fluid and preparation method thereof
  • Preserving fluid and preparation method thereof

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preparation example Construction

[0031] The present invention also provides a method for preparing the above preservation solution, comprising: mixing 0.1-10g / L buffer salt, 100-300g / L antibody stabilizer, 0.1-1g / L antiseptic and bacteriostatic agent, 2-12g / L karaage The glue is mixed with water, heated and dissolved to obtain a preservation solution.

[0032] According to the present invention, it is preferred to add 1 to 100 g / L of a thickener other than carrageenan. Wherein, the buffer salt, antibody stabilizer, antiseptic and antibacterial agent, carrageenan, water and thickeners other than carrageenan are all the same as above, and will not be repeated here.

[0033] In the present invention, it is preferred to mix 0.1-10g / L buffer salt, 100-300g / L antibody stabilizer, 0.1-1g / L antiseptic and antibacterial agent, 2-12g / L carrageenan and part of water first, and the mixing The method is preferably ultrasonic or stirring, heating and dissolving, and then adding the remaining water to constant volume to ob...

Embodiment 1

[0039] Add 700ml sterilized water to a 1000ml Erlenmeyer flask, then add 1.8gNa 2 HPO 4 12H 2 O, 0.22gKH 2 PO 4 , 3gNaCl, 0.3gKCl, 50gBSA, 150g glycerin, 0.3g sodium azide and 0.5g Proclin300, ultrasonically mixed, filtered, heated in a water bath at 40°C, 3g polyethylene glycol 12000, 6g carrageenan kappa type, 1g alginic acid were added in batches Dissolve sodium and 1g of polyvinyl alcohol until completely dissolved, then add water to make up to 1000ml to obtain a preservation solution.

[0040] Uniformly disperse the C-reactive protein-coated magnetic nanoparticles into the preservation solution.

[0041] Performance test: Use a pipette gun to accurately pipette 100 μl of 0.1mol / L phosphate buffer (PB6.5) with a pH value of 6.5 into a 1.5mL centrifuge tube, and add 10 μl of 0.1 μg / ml CRP antigen or 1 μg / ml CRP antigen, 20 μl of alkaline phosphatase-labeled antibody and 100 μl of preservation solution dispersed with magnetic nanoparticles, and then use a pipette to dis...

Embodiment 2

[0049] Add 700ml sterilized water to a 1000ml Erlenmeyer flask, then add 2.3gMOPS, 0.19gNaOH, 2.2gNaCl, 1.3gKCl, 60g calf serum, 140g glycerol, 0.3g sodium azide and 0.2g chloramphenicol, and mix by ultrasonic , filtered, heated in a water bath at 40°C, 5g of polyvinylpyrrolidone 360, 3g of carrageenan λ type, 3g of carrageenan ι type, 1g of gellan gum and 1g of polyvinyl alcohol were added in batches until they were completely dissolved, and then water was added to set the volume to 1000ml to obtain Preservation solution.

[0050] Will β 2 - The MG-coated magnetic nanoparticles are evenly dispersed into the preservation solution.

[0051] Performance test: Use a pipette gun to accurately pipette 100μl, 0.1mol / LPB6.5 to 1.5mL centrifuge tube, add 10μl 0.05μg / mlβ to it in turn 2 - MG antigen or 0.5 μg / ml β 2 - MG antigen, 20 μl of alkaline phosphatase-labeled antibody and 100 μl of preservation solution dispersed with magnetic nanoparticles, then use a pipette to disperse th...

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Abstract

The invention provides a preserving fluid and its preparation method. The preserving fluid comprises 0.1-10 g / L of buffer salt, 100-300 g / L of an antibody stabilizer, 0.1-1 g / L of an anticorrosive bacteriostatic agent and 2-12 g / L of carragheenan and water. In comparison with the prior art, the invention has the following advantages: the buffer salt, antibody stabilizer and anticorrosive bacteriostatic agent in the preserving fluid maintain stability of magnetic micro-particles or magnetic nano-particles and antibody in the whole system; and viscosity of the carragheenan can decrease exponentially with rising of temperature. Solubility of the carragheenan increases with rising of temperature, molecular dissociation is intensified, electrostatic attraction between half-esterfied sulfate radicals is weakened, and molecular entanglement decreases. Then, viscosity is lowered. But viscosity increases when temperature is reduced. At 30 DEG C, molecules are gradually entangled to form a net structure so as to sharply increase viscosity. Thus, the preserving fluid is in a viscous or gelatinous state at the temperature of 2-20 DEG C but is in a fluid state at the operating temperature of 35-38 DEG C.

Description

technical field [0001] The invention belongs to the field of chemiluminescence immunoassay, in particular to a preservation solution and a preparation method thereof. Background technique [0002] Chemiluminescence immunoassay (Chemiluminescenceimmunoassay, CLIA) is to combine the chemiluminescence system with the immune reaction, label the antibody or antigen with chemiluminescence-related substances, react with the antigen or antibody to be tested, and separate the free chemiluminescence markers, Other related substances added to the chemiluminescence system produce chemiluminescence for quantitative or qualitative detection of antigens or antibodies. Since its development in the 1970s, chemiluminescence immunoassay has become a mature and advanced ultra-trace active substance detection technology. Due to its advantages of high sensitivity, strong specificity, wide detection range, simple operation, high degree of automation, good reagent stability, and very little pollut...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/553
Inventor 冯瑾
Owner SINOCARE
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