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Double antibody complex retinol-binding protein assay kit

A detection kit and protein binding technology, applied in the field of medical immunology in vitro diagnosis, can solve the problems of insufficient titer, insufficient antibody purity, insufficient sensitivity, etc., and achieve accurate and reliable measurement results, improve accuracy, and save costs. Effect

Inactive Publication Date: 2010-09-15
浙江康特生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of purity or titer of the antibody in the current market kits for the determination of retinol-binding protein, at the same time, only one antibody is used in the reagent preparation process, resulting in good specificity and insufficient sensitivity of the kit, or high sensitivity but less specific

Method used

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  • Double antibody complex retinol-binding protein assay kit
  • Double antibody complex retinol-binding protein assay kit
  • Double antibody complex retinol-binding protein assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The composition of the assay kit is as follows:

[0029] R1:

[0030] pH7.4 phosphate buffer 45mmol / L

[0031] Polyethylene glycol 6000 78mmol / L

[0032] Disodium EDTA 10.2mmol / L

[0033] R2:

[0034] Mouse anti-human monoclonal antibody 2ml / L

[0035] Rabbit anti-human polyclonal antibody 3ml / L

[0036] pH7.4 phosphate buffer 45mmlo / L

[0037] Disodium EDTA 10.2mmlo / L

[0038] Human Serum Matrix Calibrators:

[0039] Concentration: 110mg / L

[0040] Sodium azide 0.2%

[0041] Tween-20 1%

[0042] Bovine Serum Albumin 1%

[0043] The retinol-binding protein assay kit described in this example is suitable for various types of automatic biochemical analyzers. Taking the Hitachi 7060 automatic biochemical analyzer as an example, its operation is as follows:

[0044]

[0045] Perform single-point calibration and multi-point calibration on the obtained results. The multi-point calibration adopts the Spline equation in nonlinear calibration, and the instrument ...

Embodiment 2

[0057] The composition of the Retinol Binding Protein Assay Kit is as follows:

[0058] R1:

[0059] pH7.6 phosphate buffer 60mmol / L

[0060] Polyethylene glycol 6000 95mmol / L

[0061] Disodium EDTA 13mmol / L

[0062] R2:

[0063] Mouse anti-human monoclonal antibody 3ml / L

[0064] Rabbit anti-human polyclonal antibody 4ml / L

[0065] pH7.6 phosphate buffer 60mmol / L

[0066] Disodium EDTA 13mmol / L

[0067] Human serum matrix calibrator:

[0068] Concentration: 110mg / L

[0069] Sodium Azide 1.2%

[0070] Tween-20 5.5%

[0071] Bovine Serum Albumin 2%

[0072] The experimental process is the same as in Example 1.

[0073] from Figure 5 It can be seen from the calibration curve of the 110mg / L RPB calibrator that after 8 concentration dilutions, the measured absorbance is basically proportional to the diluted concentration, and the multi-point calibration curve is a straight line. and Figure 4 The absorbance value of the calibrator concentration of 110mg / L during si...

Embodiment 3

[0084] The composition of the Retinol Binding Protein Assay Kit is as follows:

[0085] R1:

[0086] pH7.2 phosphate buffer 35mmol / L

[0087] Polyethylene glycol 6000 60mmol / L

[0088] Disodium EDTA 6mmol / L

[0089] R2:

[0090] Mouse anti-human monoclonal antibody 4ml / L

[0091] Rabbit anti-human polyclonal antibody 5ml / L

[0092] pH7.2 phosphate buffer 35mmol / L

[0093] Disodium EDTA 6mmol / L

[0094] Human serum matrix calibrator:

[0095] Concentration: 110mg / L

[0096] Sodium Azide 2.2%

[0097] Tween-20 10%

[0098] Bovine Serum Albumin 3%

[0099] The experimental process is the same as in Example 1.

[0100] from Figure 8 It can be seen from the calibration curve of the 110mg / L RPB calibrator that after 8 concentration dilutions, the measured absorbance is basically proportional to the diluted concentration, and the multi-point calibration curve is a straight line. and Figure 7 The absorbance values ​​of the calibrator concentration of 110mg / L during si...

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Abstract

A retinol-binding protein assay kit in the market at present has good specificity and insufficient sensitivity or has high sensitivity and poor specificity because the purity of antibodies is insufficient or the potency cannot meet the requirement. The invention provides a double antibody complex retinol-binding protein assay kit, which consists of three parts, namely, a reagent R1, a reagent R2 and calibration materials. The reagent R1 is a phosphate buffer system which consists of phosphate buffer solution with the pH of 7.2 to 7.6, polyethylene glycol 6000-8000 and ethylene diamine tetraacetic acid; the reagent R2 is antibody solution which consists of mouse anti-human monoclonal antibody, rabbit anti-human polyclonal antibody, phosphate buffer solution with the pH of 7.2 to 7.6 and the ethylene diamine tetraacetic acid. By adopting a complex antibody of the polyclonal antibody and monoclonal antibody, the sensitivity and high linearity are guaranteed, and the accuracy of the measured result is greatly improved.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis of medical immunity, in particular to a kit for detecting retinol binding protein in serum or urine. Background technique [0002] Retinol-binding protein (RBP) is a transporter of retinol (vitamin A) in the blood. In 1961, Berggard discovered in immunoelectrophoresis in α 2 -Proteins whose globulin domains form a long precipitation line, formerly known as long alpha 2 -globulin. In the following decades, people conducted a comprehensive study on the molecular structure and biological characteristics of this protein, and found that this protein is widely distributed in normal human body fluids, and belongs to the α 1 - A globulin that has the function of transporting retinol from hepatocytes to surrounding tissues. It is now believed that RBP in the blood mainly exists in the form of complexes bound by retinol and prealbumin. When retinol in the complex binds to target cells, RBP is separated...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531
Inventor 尹鹏
Owner 浙江康特生物科技有限公司
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