Kit for detecting apolipoprotein A5 for diagnosis of hypertriglyceridemia

A technology for hypertriglyceridemia and apolipoprotein, which is applied in disease diagnosis, biological testing, measuring devices, etc. It can solve the problems of sensitivity and accuracy limitation, narrow linear range of ApoA5, ​​etc., and improve sensitivity and accuracy , Improving the ability of antibody capture and detecting the effect of strong specificity

Inactive Publication Date: 2014-11-26
WUHAN J H BIO TECH
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

However, this method has a narrow linear range for the determination of ApoA5, ​​so its sensitivity and accuracy are limited.

Method used

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  • Kit for detecting apolipoprotein A5 for diagnosis of hypertriglyceridemia
  • Kit for detecting apolipoprotein A5 for diagnosis of hypertriglyceridemia
  • Kit for detecting apolipoprotein A5 for diagnosis of hypertriglyceridemia

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Experimental program
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Embodiment Construction

[0027] The production method of hypertriglyceridemia (HTG) diagnostic kit:

[0028] 1. Preparation of ApoA5 monoclonal antibody pairs

[0029] Purified ApoA5 was used as antigen to immunize BALB / c mice respectively, for the second time at an interval of 21 days, and once at intervals of 10 days thereafter, for a total of 3-4 times, and then the spleen cells of the immunized mice were taken out, and the cells were immunized with PEG. The spleen cells were fused with the mouse myeloma cell line SP2 / 0, the hybridoma cells were screened on a 96-well cell culture plate using HAT selective medium, and the anti-ApoA5 cell line was identified by ELISA, and cloned three times to obtain For cell lines that stably secrete highly specific monoclonal antibodies, use the ELISA inhibition method on the same antigen-coated plate, that is, add cell culture supernatant to the reference group, add cell culture supernatant and antigen mixture to the inhibition group, and compare the OD values ​​o...

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Abstract

The invention relates to a kit for detecting an apolipoprotein A5 (ApoA5) for diagnosis of hypertriglyceridemia. The kit contains two unlabeled anti-ApoA5 monoclonal antibodies, an enzyme-labeled anti-ApoA5 monoclonal antibody, a blocking solution, an enzyme substrate solution, a coating buffer solution, a stop solution, a washing solution, and an ELISA plate; the two unlabeled anti-ApoA5 monoclonal antibodies coat the ELISA plate and are used as capture antibodies; the enzyme-labeled anti-ApoA5 monoclonal antibody is used as a detection antibody; the coating buffer solution is a carbonate buffer solution with the concentration of 0.05 mol/L and the pH of 9.6, wherein 1 L of the solution contains 1.59 g of Na2CO3 and 2.93 g of NaHCO3; the blocking solution is a bovine serum albumin with the mass concentration of 1%; the enzyme substrate solution is a phosphate buffer solution having the concentration of 0.1 mol/L and the pH of 6.0 and containing 3,3',5',5-tetramethylbenzidine with the concentration of 60 [mu]g/mL and H2O2 with the mass concentration of 0.05%; the stop solution is an H2SO4 solution with the concentration of 1 mol/L. The two monoclonal antibodies coat the ELISA plate to capture ApoA5 in human blood, the antibody capture ability is improved, the determination sensitivity and accuracy and linear range are improved, and test and clinical requirements are reached.

Description

technical field [0001] The invention relates to apolipoprotein A5 (ApoA5) as a biochemical index for the diagnosis of hypertriglyceridemia (HTG), the preparation technology of anti-ApoA5 antibody pairs, the technology of enzyme-labeled chromogenic immune reaction and the use of ApoA5 antibodies to diagnose high triglycerides The technical field of hypertriglyceridemia (HTG), in particular to a kit for quantitatively detecting ApoA5 for diagnosing hypertriglyceridemia (HTG). Background technique [0002] There are many items for clinical detection of blood lipids. The basic items commonly checked by hospitals include total cholesterol (TC), triglyceride (TG), apolipoprotein (Apo) B, Apo AI, lipoprotein (a) [Lp(a )]Wait. Survey data studies have shown that high triglycerides, high cholesterol, low high-density lipoprotein-C (HDL-C) and high low-density lipoprotein-C (LDL-C) together constitute the risk factors for coronary heart disease. Abnormal apolipoprotein (Apo) is clos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/92
CPCG01N33/92G01N33/577G01N2800/32G01N2800/324
Inventor 钱金宏廖园园廖伟
Owner WUHAN J H BIO TECH
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