Long-acting ELISA plate stabilizing agent

A technology of enzyme label plate and stabilizer, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of Tween 20 color depth, certain influence of the test, and peculiar smell, and achieve simple formula and easy-to-obtain components , cost reduction effect

Active Publication Date: 2013-12-25
河北渤腾医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Chinese patent CN201110162393.2 discloses a microporous plate sealing stabilizer, which includes proteins, sugars, organic polymers, nonionic surfactants, preservatives and buffers, which can effectively prolong the validity period of microporous plates; Ionic surfactants are widely used in the field of biological sciences, but they still have some biological toxicity. For example, Tween surfactants have hemolytic effect, and Tween 20 has deep color and odor
Chinese patent CN201210116851.3 discloses a blocking solution in ELISA in vitro diagnostic reagents, which includes buffer, bovine serum albumin, trehalose, protein stabilizer, L-lysine, Proclin300 and sodium hydroxide or hydrochloric acid, wherein Although sodium hydroxide or hydrochloric acid is only used as an acid-base regulator, the amount added is small, but because it is a strong acid or strong base, the cations and anions produced after ionization will still have a certain impact on the test

Method used

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  • Long-acting ELISA plate stabilizing agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Recombinant Human Serum Albumin 0.5g

[0036] Dextran 10g

[0037] Glycerin 10ml

[0038] ProClin200 0.02%

[0039] Phosphate buffer 100mM, pH7.4

[0040] EDTA Disodium Salt 5mmol

[0041] Add water to make up to 100ml.

[0042] Add 100 μL of the above-mentioned stabilizer to the microtiter plate coated with the alpha-fetoprotein antibody, and use no stabilizer as the control group.

[0043] Method of use: Add 100 μL of the above stabilizer to the microplate, incubate at 37°C for 1 hour, place the microplate in a vacuum drying oven, dry it in vacuum, and package it.

[0044] Place the two kinds of microplates in a 37-degree environment for 2, 4, 6, 8, 10, 12, and 14 days, and use the 0-day microplate as a control to detect and calculate the enzyme-linked immunoassay. Residual activity of the target.

[0045] As can be seen from the data in Table 1, the enzyme label plate without stabilizer loses its activity soon, while the activity of the enzyme label plate of E...

Embodiment 2

[0049] Casein hydrolyzate 2.5g

[0050] Inulin 15g

[0051] Sorbitol 2g

[0052] ProClin300 0.05%

[0053] Phosphate buffer 100mM pH7.4

[0054] EDTA Disodium Salt 5mmol

[0055] Add water to make up to 100ml.

[0056] Add 100 μL of the above-mentioned stabilizer to the microtiter plate coated with the alpha-fetoprotein antibody, and use no stabilizer as the control group.

[0057] Method of use: Add 100 μL of the above stabilizer to the microplate, incubate at 37°C for 1 hour, place the microplate in a vacuum drying oven, dry it in vacuum, and package it.

[0058] Place the two kinds of microplates in a 37-degree environment for 2, 4, 6, 8, 10, 12, and 14 days, and use the 0-day microplate as a control to detect and calculate the enzyme-linked immunoassay. Residual activity of the target.

[0059]As can be seen from the data in Table 2, the enzyme label plate that does not add stabilizer loses activity very soon, and adds the enzyme label plate of embodiment 2 as time ...

Embodiment 3

[0063] Bovine Serum Albumin 5g

[0064] Inulin 1g

[0065] Mannitol 1g

[0066] ProClin300 0.1%

[0067] Phosphate buffer 100mM pH7.4

[0068] EDTA Disodium Salt 5mmol

[0069] Add water to make up to 100ml.

[0070] Add 100 μL of the above-mentioned stabilizer to the microtiter plate coated with the alpha-fetoprotein antibody, and use no stabilizer as the control group.

[0071] Method of use: Add 100 μL of the above stabilizer to the microplate, incubate at 37°C for 1 hour, place the microplate in a vacuum drying oven, dry it in vacuum, and package it.

[0072] Place the two kinds of microplates in a 37-degree environment for 2, 4, 6, 8, 10, 12, and 14 days, and use the 0-day microplate as a control to detect and calculate the enzyme-linked immunoassay. Residual activity of the target.

[0073] As can be seen from the data in Table 3, the enzyme label plate that does not add stabilizer loses activity very soon, and adds the enzyme label plate of embodiment 3 as time g...

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Abstract

The invention relates to a long-acting ELISA (Enzyme-linked Immunosorbent Assay) plate stabilizing agent for keeping protein activity in an ELISA plate. The agent comprises 0.1-5g protein, 1-15g sugar, 0.1-10g organic matter, 0.01-0.1%(V/V) of preservative, and 10-200mmol buffer solution salt per 100ml water, and a pH (Potential of Hydrogen) value is 7.2-7.4. The stabilizing agent adopts a technical route integrating a closing process and a stabilization process, and can effectively close excessive sites in the ELISA plate, and some active ingredients in the stabilizing agent can ensure continuous stabilization of the ELISA plate. The agent is simple in formula, can stabilize the ELISA plate, and does not produce any influence on a result simultaneously. The ingredients of the agent are easy to obtain, and the formula is simple, so that the cost of the stabilizing agent is lowered.

Description

technical field [0001] The invention relates to a microplate plate stabilizer for maintaining protein activity in the microplate plate. Background technique [0002] Immunoassay technology has been widely used in today's clinical testing and life science research, with a huge number of related products and broad market prospects. Immunoassay techniques include enzyme-linked immunoassay technology, chemiluminescence immunoassay technology, time-resolved fluorescence immunoassay technology, etc. Most of the solid phase carriers used in these detection technologies are enzyme-labeled plates. [0003] As one of the key components of the immunoassay kit, the enzyme plate must not only have certain sensitivity and specificity, but also its repeatability and stability are very important. Good stability is an important symbol of its data reliability. However, the stability of some enzymes and protein markers is very poor. When the antigen or antibody is coated on the microtiter p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 程华闫静辉吴萌董超李亚璞
Owner 河北渤腾医药技术有限公司
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