Method and reagent box for expanding immune detecting measurable range

A technology of immune detection and measurement range, which is applied in the detection field and can solve problems such as tedious work, narrow detection range, and increased detection cost

Active Publication Date: 2012-11-14
SHANGHAI TELLGEN LIFE SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the detection method is determined and the antibody (or antigen) raw material is selected, the measurement range is as large as it is determined, and the amount of raw material used can only change the measurement range within a small range; different immunoassay methods have limitations on the measurable range of reagents Either the sensitivity is low or the detection range is narrow. Even if some advanced technology platforms (such as ECL, FTIR, etc.) are used, the sensitivity and detection range can only be slightly increased.
[0003] The concentration of the target detection substance in clinical samples (such as serum samples) often exceeds the high value detection range of the kit. For example, the general measurement range of the alpha-fetoprotein (AFP) kit is 0 to 2000ng / ml, but the serum samples of AFP in the population The value distribution range is 0 to 2000,000ng / ml, and many patient serum samples are much higher than 2000ng / ml, distributed between 10,000-100,000ng / ml, in this case, for the one-step method (while reacting at the same time Addition of solid-phase carriers with immobilized capture antibodies or antigens, serum samples, secondary antibody markers and signal substances, without washing process) often report false-negative results. The upper limit of the detection range of the kit and cannot give a specific value
In view of this situation, the existing kits generally use multiple dilutions and repeated measurements to give specific values, which brings a lot of cumbersome work to the staff and increases the cost of testing, which is a bottleneck of the existing immunoassay kits

Method used

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  • Method and reagent box for expanding immune detecting measurable range
  • Method and reagent box for expanding immune detecting measurable range
  • Method and reagent box for expanding immune detecting measurable range

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] Quantitative detection of alpha-fetoprotein (AFP) in human serum

[0125] Experimental Materials

[0126] The primary and secondary antibodies against AFP were purchased from Shanghai Second Military Medical University,

[0127] The pure AFP antigen was purchased from Biodesign,

[0128] Beads were purchased from Luminex,

[0129] Conventional reagents are commercially available products.

[0130] experimental method

[0131] 1. Preparation

[0132] 1.1 The primary antibody and AFP antigen are pre-removed of amino-containing small molecules and protein (dialysis or chromatography column), and their concentration is measured.

[0133] 1.2 In a 1.5ml polypropylene centrifuge tube, accurately weigh about 5mg of N-hydroxysμLfosuccini-mide (NHSS) for use (moisture-proof).

[0134] 1.3 In a 1.5ml polypropylene centrifuge tube, accurately weigh about 5mg of N-ethyl-N’(3-dimethylainopropyl)-carbodiimide (EDC) for use (moisture-proof).

[0135] 2.beads activation

[0136] 2.1 Respectively take N...

Embodiment 2

[0197] Quantitative detection of alpha-fetoprotein (AFP) in human serum (Cytometric Bead Array (CBA))

[0198] Experimental Materials

[0199] The primary and secondary antibodies against AFP were purchased from Shanghai Second Military Medical University,

[0200] The pure AFP antigen was purchased from Biodesign,

[0201] Beads purchased from BD

[0202] BD FACSCalibur,

[0203] Conventional reagents are commercially available products.

[0204] experimental method

[0205] 1. Preparation

[0206] 1.1 The primary antibody and AFP antigen are pre-removed of amino-containing small molecules and impurities (dialysis or chromatography column), and their concentration is measured.

[0207] 1.2 In a 1.5ml polypropylene centrifuge tube, accurately weigh about 5mg of N-hydroxysμLfosuccini-mide (NHSS) for use (moisture-proof).

[0208] 1.3 In a 1.5ml polypropylene centrifuge tube, accurately weigh about 5mg of N-ethyl-N’(3-dimethylainopropyl)-carbodiimide (EDC) for use (moisture-proof).

[0209] 2.be...

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PUM

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Abstract

The invention discloses a method and a corresponding kit to expand the detectable range of objects to be detected in immunity analysis.

Description

Technical field [0001] The invention relates to the technical field of in vitro immunoassays, in particular to a detection method, which can expand the measurable range of a target analyte. Background technique [0002] In an immunoquantitative detection test, the measurable range is affected by the detection method, the nature of the antibody (or antigen) and the amount of antibody (or antigen) used. If the detection method is determined and the antibody (or antigen) raw material is selected, the measurement range is too large to be determined, and the amount of raw material used can only change the measurement range within a small range; different immunoassay methods have limitations on the measurable range of reagents Either the sensitivity is low, or the detection range is narrow. Even if some advanced technology platforms (such as ECL, FTIR, etc.) are used, the sensitivity and detection range can only be increased by a small margin. [0003] The concentration of the target te...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543
Inventor 姚见儿罗朝领
Owner SHANGHAI TELLGEN LIFE SCI CO LTD
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