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Mass spectrometric detection method for ca-dependent secretory protein 1 in human serum

A secreted protein, mass spectrometry detection technology, applied in the field of mass spectrometry detection of calcium-dependent secreted protein 1 in human serum, can solve the problems of laborious, insufficient sensitivity, time-consuming, etc., and achieve the effect of easy detection, high repeatability, and simple operation

Inactive Publication Date: 2012-10-31
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Description
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Problems solved by technology

At present, the main detection methods of caps1 include western blotting and enzyme-linked immunosorbent assay. Their disadvantages are time-consuming, labor-intensive, and insufficient sensitivity, so they are not suitable for the screening of large-scale samples.

Method used

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  • Mass spectrometric detection method for ca-dependent secretory protein 1 in human serum

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Experimental program
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Embodiment

[0012] 1. Main reagents:

[0013] Weak cation exchange magnetic beads MB-WCX kit (Bruker Daltonik GmbH, Germany),

[0014] Immunometal affinity copper ion magnetic beads MB-IMAC Cu kit (Bruker Daltonik GmbH, Germany)

[0015] α-Cyano-4-hydroxycinnamic acid α-Cyano-4-hydroxycinnamic acid (HCCA) (sigma, USA)

[0016] Acetone (sigma, USA)

[0017] Ethanol (sigma, USA)

[0018] Anchor Chip TM Target (Bruker Daltonik GmbH, Germany)

[0019] 2. Consumables and instruments:

[0020] Vacuum blood collection tube (BD Vacutainer TM ,United States)

[0021] Low temperature refrigerator (SANYO, Japan)

[0022] Vortex mixer (H101, Shanghai Kanghe Photoelectric Instrument Co., Ltd.)

[0023] Constant temperature high-speed centrifuge (Z400K, HERMLE, Germany)

[0024] Magnetic bead separator (Bruker Daltonik GmbH, Germany)

[0025] Autoflex II matrix-assisted laser dissociation ion time-of-flight mass spectrometry (Bruker Daltonik GmbH, Germany)

[0026] LC / MS / MS instrument (Appl...

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Abstract

The invention relates to a mass spectrometric detection method for ca-dependent secretory protein 1 in human serum, which is characterized in that the mass spectrometric detection method comprises the following steps: taking human serum and using magnetic bead attachment protein for obtaining magnetic bead enrichment liquid; adding the magnetic bead enrichment liquid into a matrix solution; carrying out the matrix-assisted laser dissociation ionization flight time mass spectrometric detection according to the following conditions: a first ion source voltage: 20kV; a second ion source voltage:18.6kV; crystal voltage: 7.6kV; pulse ion time: 320ns; polarity: positive; matrix inhibition mode: gate control; gate control strength: maximum: suppression high limit: 600Da; detector: 1400-1500 V; sample rate: 1.00Gs / s; electrical acquisition: enhancing, 100mV; real-time fluency: high; laser frequency: 20Hz; laser attenuation: closing 73% and ranging 20%; mass to charge ratio is 5336 plus or minus 1; and the peak flowing is the ca-dependent secretory protein 1. The method has the advantages of short time and high sensitivity.

Description

technical field [0001] The present invention relates to a mass spectrometric detection method for calcium-dependent secreted protein 1 in human serum, in particular to a method using magnetic bead enrichment combined with matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (matrix-assisted laserdesorption / ionisation time-of-flight mass spectrometry) The invention discloses a method for screening calcium-dependent secretory protein 1 (caps1) from human serum by spectrometry, MALDI-TOF MS, and belongs to the technical field of protein detection methods. Background technique [0002] Calcium-dependent secretion activator 1 (CAPS-1) is a calcium-binding protein that can promote the transport of neurotransmitters and neuropeptide vesicles to the cell membrane for further release of secretory granules. At present, the main detection methods of caps1 include western blotting and enzyme-linked immunosorbent assay. Their disadvantages are time-consuming, labo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543
Inventor 沈锡中薛如意刘韬韬董玲
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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