Double antibody latex enhanced retinol binding protein detection kit

A protein-binding and latex-enhanced technology, applied in biological testing, material inspection products, etc., can solve the problems of insufficient sensitivity and inability to simultaneously RBP, etc., to achieve the effect of ensuring accuracy

Active Publication Date: 2012-08-08
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Although there are a variety of kits for immunoturbidimetric methods on the market, due to insufficient sensitivity, they all have the disadvantage that the same reagent cannot simultaneously detect RBP in serum samples and urine samples, and almost all kits are used for RBP in serum samples. detection

Method used

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  • Double antibody latex enhanced retinol binding protein detection kit
  • Double antibody latex enhanced retinol binding protein detection kit
  • Double antibody latex enhanced retinol binding protein detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1: RBP kit 1 composition

[0061] Reagent 1 component:

[0062] Buffer MOPS 0.15mol / L pH=6.5,

[0063] PEG6000 10%,

[0064] NaN 3 0.1%,

[0065] EDTA 100mmol / L,

[0066] Reagent 2 components:

[0067] Buffer MOPS 0.15mol / L pH=8.5,

[0068] Monoclonal antibody A 0.45mg / ml (source: mouse anti-human, antigen: retinol binding protein extracted from human urine, potency: greater than 1:5000),

[0069] Latex particle A (for monoclonal antibody A coating) 104nm 0.35% (manufacturer: JSR carboxyl group amount: 0.238meq / g, concentration: 5%),

[0070] Monoclonal antibody B 0.5mg / ml (source: mouse anti-human, antigen: retinol binding protein extracted from human urine, potency: greater than 1:1000),

[0071] Latex particle B (for monoclonal antibody B coating) 77nm 0.5% (manufacturer: JSR, carboxyl content: 0.274meq / g, concentration: 5%),

[0072] Calibrators: Serum calibrators with concentrations of 130, 100, 50, 25, 12.5, 0mg / L,

[0073] Calibrator: urine ...

Embodiment 2

[0074] Embodiment 2: The sensitivity performance evaluation of the kit 1 of the present application

[0075] Instrument: Olympus AU400

[0076] Table 1: Parameters:

[0077]

[0078] Urine calibrator 4, 2, 1, 0.5, 0.25, 0mg / L,

[0079] Control reagent:

[0080] A well-known foreign brand A:

[0081] Its main components:

[0082] Reagent R1: GOOD’S buffer,

[0083] Reagent R2: latex-coated rabbit anti-human RBP antibody IgG,

[0084] Detection principle:

[0085] Latex particles coated with multiple anti-RBP antibodies bind to the RBP antigen in the sample to form an immune complex, and the turbidity change is detected at 570nm, and the degree of change is proportional to the RBP content in the sample.

[0086] Urine calibrator 5, 2.5, 1.25, 0.625, 0.31, 0mg / L,

[0087] Sensitivity evaluation method:

[0088] Dilute the sample prepared from pure RBP so that a certain concentration is close to the sensitivity limit, repeat 10 times to detect the water blank and sampl...

Embodiment 3

[0108] Example 3: Linear performance evaluation of kit 1 of the present application

[0109] Instrument: Olympus 400

[0110] Table 8: Parameters:

[0111]

[0112] Serum calibrator: the concentrations are 130, 100, 50, 25, 12.5, 0mg / L,

[0113] Linear evaluation method:

[0114] Serum samples were diluted in a gradient of 1 / 10, 1 / 40, 1 / 20, 1 / 10, 2 / 10, 3 / 10, 4 / 10, 5 / 10, 6 / 10, 7 / 10, 8 / 10, 9 / 10, 1, each concentration was tested 3 times. Take the dilution ratio as the X-axis, and use the measured value as the Y-axis to draw a graph. The results are shown in figure 1 .

[0115] Table 9: Linearity detection data:

[0116]

[0117] Linear regression plotting was performed on the data, and the results are shown in the appendix figure 1 Evaluation criteria: The deviation between the detection and theoretical value should be within ±10%, which is acceptable.

[0118] Conclusion: when the sample concentration is greater than 130 mg / L, prozone will appear in the detection; th...

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Abstract

The invention relates to a double antibody latex enhanced retinol binding protein detection kit. More specifically, the invention discloses a double antibody coating latex enhanced immunoturbidimetry kit for detecting retinol binding protein. The kit contains a reagent 1, a reagent 2 and a calibrator. Paring monoclonal antibody A and B are respectively coated on latex particles. Coated antibody is bonded with RBP to be detected, and a plurality of bonders are aggregated together to form detectable turbidity change. The detection kit provided by the invention has high sensitivity, can be used to detect urine samples and serum samples and can be used as nutritive index to detect RBP in serum. Simultaneously, through the detection of RBP in urine, the kit is sensitive to the damage degree of renal proximal tubule.

Description

technical field [0001] The invention relates to a double-antibody latex-enhanced detection kit for retinol-binding protein. More specifically, the invention discloses a double-antibody-coated latex-enhanced immune transmission turbidimetric kit for detecting retinol-binding protein. Includes Reagent 1 and Reagent 2 and calibrator. Background technique [0002] Retinol binding protein (RBP) consists of a polypeptide chain and a small portion of carbohydrates, does not contain neutral sugar and hexosamine, and consists of 182 amino acids; the molecular weight is 21KD; the isoelectric point is pH4.4- 4.8, half-life 3-12h. [0003] The RBP family is divided into five classes, including secretory RBPs, intracellular RBPs, nuclear receptors for retinoic acid, visual tissue-specific extracellular RBPs, and visual tissue-specific intracellular RBPs. [0004] RBP is widely distributed in normal body fluids and has the function of transporting retinol from liver cells to surrounding...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
Inventor 李彦超高爱民刘希
Owner BEIJING STRONG BIOTECH INC
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