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25-hydroxyvitamin D analysis method based on combination of immune purification and mass spectrometric detection

A hydroxyvitamin and analysis method technology, applied in the field of 25-hydroxyvitamin D analysis based on immunopurification combined with mass spectrometry detection, can solve problems affecting quantification and limiting clinical applications, cross-interference, etc., achieve strong specificity and selectivity, and improve Use efficiency, high flux effect

Active Publication Date: 2020-07-03
SUZHOU SYM BIO LIFESCI CO LTD
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  • Claims
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Problems solved by technology

However, both 25-(OH)D2 and 25-(OH)D3 are generally captured by antibodies, so the immunoassay method detects the level of total 25-(OH)D, and is also susceptible to other 25-(OH)D structures Cross-interference of similar analytes, affecting quantitation and limiting clinical application

Method used

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  • 25-hydroxyvitamin D analysis method based on combination of immune purification and mass spectrometric detection
  • 25-hydroxyvitamin D analysis method based on combination of immune purification and mass spectrometric detection
  • 25-hydroxyvitamin D analysis method based on combination of immune purification and mass spectrometric detection

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Embodiment

[0036] refer to figure 1 , this embodiment provides a method for analyzing 25-hydroxyvitamin D based on immunopurification combined with mass spectrometry detection, comprising the following steps:

[0037] Step 1, prepare biotin-labeled antibody:

[0038] Take 1 mg of 25-(OH)D antibody and dialyze it with phosphate buffer (pH 8.1) at 4°C or room temperature for no less than 16-24 hours; add 0.05 mg of biotin to the dialyzed antibody, and then add phosphate buffer ( pH 8.1) to make the final antibody concentration 1mg / mL, mix for 5 minutes; place the mixture in a constant temperature incubator at 25±1°C, let it stand for 120±10min, and finally use phosphate buffer (pH 7.4) at 4°C or Dialyze the biotinylated antibody for 16-24 hours at room temperature.

[0039] Step 2, prepare biotin-labeled antibody-coated magnetic beads:

[0040] Shake the streptavidin magnetic beads well, then absorb 20 mg of magnetic beads (volume x μL), wash twice with magnetic bead washing solution no...

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Abstract

The invention discloses a 25-hydroxyvitamin D analysis method based on combination of immune purification and mass spectrum detection. The method mainly comprises steps of preparing a biotin-labeled antibody, preparing a biotin-labeled antibody-coated magnetic bead, enriching and purifying a sample and carrying out mass spectrum detection. The analysis method is advantaged in that high specificityof antibody affinity is utilized, 25-(OH) D can be specifically enriched and purified from a complex biological sample, moreover, respective accurate detection of 25-(OH) D2 and 25-(OH) D3 is realized, the antibody coupled magnetic bead enrichment method is combined with mass spectrum detection, so high flux, sensitivity and accurate quantification are realized, moreover, use efficiency of a liquid phase mass spectrum system is greatly improved.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a 25-hydroxyvitamin D analysis method based on immunopurification combined with mass spectrometry detection. Background technique [0002] 25-hydroxyvitamin D (25-(OH)D), including 25-hydroxyvitamin D2 (25-(OH)D2) and 25-hydroxyvitamin D3 (25-(OH)D3), is the main presence of vitamin D in the body form. The level of serum 25-(OH)D can reflect the storage level of vitamin D in the human body, and it is related to the clinical symptoms of vitamin D deficiency. The detection methods of 25-(OH)D2 and 25-(OH)D3 in biological samples (such as whole blood, serum, urine or saliva) mainly include liquid chromatography-mass spectrometry and immunoassay. [0003] Before detection by liquid chromatography mass spectrometry, 25-(OH)D2 and 25-(OH)D3 need to be enriched and separated from biological samples. Traditional sample processing methods mainly include precipitation, liqui...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/08G01N30/14
CPCG01N30/02G01N30/06G01N30/08G01N30/14G01N2030/067
Inventor 姜宽祝长城许海峰
Owner SUZHOU SYM BIO LIFESCI CO LTD
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