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Liver-type fatty acid binding protein detection reagent box

A technology for fatty acid binding and protein detection, which is applied in biological testing, material inspection products, and analysis through chemical reactions of materials, etc. It can solve the problem of low sensitivity of detection of liver-type fatty acid binding protein

Inactive Publication Date: 2018-02-02
NANTONG EGENS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved by the present invention is the defect that the sensitivity of detecting liver-type fatty acid binding protein is not high in the prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The present embodiment provides a kind of method for preparing enzyme conjugate, comprises the following steps:

[0036] 1. Store the L-FABP-labeled antibody at a concentration of 2mg / mL; add 0.5M TCEP solution to the solution to make the final concentration 1.25mM, mix immediately, and let stand at 25°C for 60 minutes.

[0037] 2. Weigh an appropriate amount of Sulfo-SMCC reagent, and use dimethylformamide DMF to prepare a solution with a concentration of 17.5 mg / mL.

[0038] 3. Add the Sulfo-SMCC solution prepared in step 2 to the alkaline phosphatase solution (AP solution, the concentration is 20mg / ml). The molar ratio of alkaline phosphatase to Sulfo-SMCC is 1:10. Leave to react for 15 minutes.

[0039] 4. Add 1M glycine solution to the AP solution obtained after the activation reaction in step 3. The molar ratio of glycine to Sulfo-SMCC reagent is 1:10, mix immediately, and stand at 25°C for 10 minutes.

[0040] 5. Immediately desalt the antibody solution after t...

Embodiment 2

[0048] The present embodiment provides a kind of method for preparing enzyme conjugate, comprises the following steps:

[0049] 1. Store the L-FABP-labeled antibody at a concentration of 2mg / mL; add 0.5M TCEP solution to the solution to make the final concentration 1.25mM, mix immediately, and let stand at 25°C for 60 minutes.

[0050] 2. Weigh an appropriate amount of Sulfo-SMCC reagent, and use dimethylformamide DMF to prepare a solution with a concentration of 17.5 mg / mL.

[0051] 3. Add the Sulfo-SMCC solution prepared in step 2 to the alkaline phosphatase solution (AP solution, the concentration is 20mg / ml), the molar ratio of alkaline phosphatase to Sulfo-SMCC is 1:15, mix immediately, and let it stand at 25°C. Leave to react for 15 minutes.

[0052] 4. Add 1M glycine solution to the AP solution obtained after the activation reaction in step 3. The molar ratio of glycine to Sulfo-SMCC reagent is 1:20, mix immediately, and stand at 25°C for 10 minutes.

[0053] 5. Immedia...

Embodiment 3

[0061] The present embodiment provides a method for magnetic bead coating, comprising the following steps:

[0062] 1. Store the L-FABP-coated antibody in 100mM MES buffer at pH 5.0 with a concentration of 1mg / mL;

[0063] 2. Take carboxyl magnetic bead solution 100 times the weight of the antibody, the particle size of carboxy magnetic beads is 3 μm, magnetically separate and discard the supernatant;

[0064] 3. Washing: re-dissolve the magnetic beads obtained in the above steps with 100 mM MES buffer solution of pH 5.0, and discard the supernatant by magnetic separation.

[0065] 4. Repeat step 3 once

[0066] 5. Add an appropriate amount of 100mM MES buffer to dissolve the magnetic beads obtained in step 4.

[0067] 6. Weigh an appropriate amount of NHS reagent and dissolve it with 100mM MES buffer at pH 5.0 to a solution with a concentration of 10mg / mL;

[0068] 7. Weigh an appropriate amount of EDC reagent and dissolve it in 100mM MES buffer with pH 5.0 to a solution w...

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PUM

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Abstract

The invention discloses a liver-type fatty acid binding protein detection reagent box. The box comprises seven storage assemblies which are used for storing L-FABP correcting products, L-FABP qualitycontrolling products, enzyme compound working liquid, magnetic bead working liquid, cleaning liquid, substrate solutions and pretreatment reagents respectively; the magnetic bead working liquid comprises carboxyl magnetic beads with marked L-FABP antibodies, and the enzyme compound working liquid comprises L-FABP antibodies marked by alkaline phosphatase. The invention further discloses a detection method for detecting liver-type fatty acid binding proteins. According to the detection reagent box, the minimum detection limit is 0.5 ng / ml, the linear range is 0.5-500 ng / ml, the detection sensibility is high, the linear range is wide, the detection time length is shortened to 15 minutes, and the detection procedures are simplified.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a detection kit for liver-type fatty acid binding protein. Background technique [0002] Acute kidney injury (Acute kidney injury, AKI) is caused by a variety of reasons, can occur in a variety of clinical situations, a complex syndrome of renal dysfunction. It can manifest as a mildly elevated blood creatinine level or as acute kidney failure. Studies have found that the occurrence of AKI is closely related to the increase in mortality. This requires doctors to diagnose early in clinical work, preferably when the glomerular filtration rate begins to decline, or when there is only tissue damage and the glomerular filtration rate is still normal, to detect and intervene in time to improve prognosis and reduce fatality rate. Serum creatinine and urine output are current diagnostic indicators for AKI, but they are affected by many factors, and their sensitivity and specificity...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/76
CPCG01N21/76G01N33/68
Inventor 王保君汤双双褚晖欧卫军顾飞
Owner NANTONG EGENS BIOTECH
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