SA-magnetic bead freeze-drying working solution, SA-magnetic bead freeze-dried product and preparation method of freeze-dried product

A working solution and freeze-drying technology, which is applied to material inspection products, measuring devices, instruments, etc., can solve the problems of unstable luminescence intensity of SA-magnetic beads freeze-dried products, short storage time of SA-magnetic beads, and inconvenient transportation. Achieve the effect of being beneficial to freeze-drying, stable luminescence value, and small effect on activity

Active Publication Date: 2019-05-07
DONGGUAN HEC MEDICAL INTELLIGENT DEVICE R&D CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a freeze-dried working solution of SA-magnetic beads freeze-dried product and its preparation method,

Method used

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  • SA-magnetic bead freeze-drying working solution, SA-magnetic bead freeze-dried product and preparation method of freeze-dried product
  • SA-magnetic bead freeze-drying working solution, SA-magnetic bead freeze-dried product and preparation method of freeze-dried product
  • SA-magnetic bead freeze-drying working solution, SA-magnetic bead freeze-dried product and preparation method of freeze-dried product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The preparation of embodiment 1 freeze-dried working solution

[0034] Prepare the following lyophilized working solutions respectively:

[0035] Freeze-dried working solution A:

[0036] TBS buffer (10 mM Tris-HCl, 0.9% NaCl, pH: 7.4), 1% glycine, 1% inulin, 1% mannitol, 1% sodium caseinate, 0.05% Tween-20, 0.02% ProClin 300.

[0037] Freeze-dried working solution B:

[0038] TBS buffer (20 mM Tris-HCl, 0.9% NaCl, pH: 7.4), 1% glycine, 1% inulin, 2% mannitol, 2% sodium caseinate, 0.05% Tween-20, 0.02% ProClin 300.

[0039] Freeze-dried working solution C:

[0040] TBS buffer (30 mM Tris-HCl, 0.9% NaCl, pH: 7.4), 1% glycine, 2% inulin, 1% mannitol, 3% sodium caseinate, 0.05% Tween-20, 0.02% ProClin 300.

[0041] Freeze-dried working solution D:

[0042] TBS buffer (20 mM Tris-HCl, 0.9% NaCl, pH: 7.4), 1% glycine, 3% inulin, 2% mannitol, 1% sodium caseinate, 0.05% Tween-20, 0.02% ProClin 300.

[0043] Freeze-dried working solution E:

[0044] TBS buffer (40 mM Tri...

Embodiment 2

[0055] Example 2 Preparation of SA-magnetic beads freeze-dried product

[0056] 1) Adsorb SA-magnetic beads through a magnetic stand, mix SA-magnetic beads with freeze-dried working solutions A, B, C, D, E, F, G, H, I, J, and adjust the content of magnetic beads to 1 % (1 g / 100 mL).

[0057] 2) Spot SA-magnetic beads onto a 0.5cm×0.5cm glass chip with an automatic spotting instrument, 100nL per spot, 7×7 square array, 49 spots in total.

[0058] 3) Freeze-drying of SA-magnetic bead lyophilized product

[0059] Pre-freezing: the chip is put into the box at 4°C, and kept for 1 hour after entering the box, so that all samples are cooled from the same starting point of 4°C to -46°C, and kept at -46°C for 1 hour, and the samples are heated to the eutectic point The temperature was maintained for 1 hour, and then the sample was cooled again and maintained at -46°C for 2 hours.

[0060] Sublimation: Control the temperature of the plate layer to maintain the temperature of the samp...

Embodiment 3

[0064] Example 3 Redissolution and Activity Detection of SA-Magnetic Beads Freeze-dried Product

[0065] 1) Activity detection after freeze-drying and reconstitution of SA-magnetic beads

[0066] Redissolve 10 kinds of SA-magnetic bead lyophilized products with 100 μL ultrapure water;

[0067] Add 2 μL of biotinylated alkaline phosphatase (Biotin-AP) (5 ng / μL) to the 10 kinds of SA-magnetic beads after reconstitution and the same batch of non-lyophilized SA-magnetic bead suspension, and incubate at 37°C for 10 minutes.

[0068] Add 200 μL of SA-magnetic bead washing buffer, absorb and wash three times, and magnetically absorb for 3 minutes each time.

[0069] Add 200 μL enzyme substrate solution, mix well, take 170 μL to a microplate plate, incubate in a microplate reader at 37°C for 5 min, measure the luminescent signal value, and the results are shown in Table 1.

[0070] Table 1 Activity analysis of SA-magnetic beads bound to biotinylated alkaline phosphatase before and a...

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Abstract

The invention discloses an SA-magnetic bead freeze-drying working solution, an SA-magnetic bead freeze-dried product and a preparation method of the freeze-dried product. The freeze-drying working solution is suitable for an alkaline phosphatase luminescence system and is prepared from a protein protectant and a buffer solution, wherein casein sodium is adopted as the protein protectant, a TBS buffer solution is adopted as the buffer solution, the TBS buffer solution is prepared from the components of 10-50 mM of Tris-HCl and 0.9% NaCl, and the pH is 7.4. The freeze-drying working solution isprepared from the casein sodium and the TBS buffer solution, so that the freeze-drying working solution is more suitable for the SA-magnetic bead freeze-dried product of an alkaline phosphatase reaction system; and a mixed solution of SA-magnetic beads and the freeze-drying working solution is subjected to sample application to the interior of a chip in micro-droplets, then after in-situ freeze-drying, the freeze-dried product is obtained, the freeze-dried product can keep good biotin combining activity after being redissolved, after the freeze-dried product generates a sandwich reaction witha biotinylated antibody and antigen and an alkaline phosphatase marked antibody, influence on activity of an enzyme catalyzed substrate is small, and the luminescence value is high and stable.

Description

technical field [0001] The invention relates to the technical field of biochemical reagents, in particular to a streptavidin-coupled magnetic bead (SA-magnetic bead) working solution, a streptavidin-coupled magnetic bead (SA-magnetic bead) freeze-dried product and Preparation method of freeze-dried product. Background technique [0002] Currently commercially available SA-magnetic beads are stored in low temperature (4-8°C) buffer. The SA-magnetic beads are suspended and absorbed into a certain volume, and can be directly used in a liquid-phase immunoreaction system based on SA-magnetic beads, biotinylated antibodies and enzyme-labeled antibodies. However, liquid-preserved SA-magnetic beads are not suitable for use in protein chips or microfluidic chips. Protein chips or microfluidic chips require pre-embedded freeze-dried solid-phase SA-magnetic beads, and SA-magnetic beads are required It can still have a certain skeleton structure. When in use, SA-magnetic beads can be ...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531
Inventor 陈立勇邓晓侠刘仁源宋建军顾志鹏刘勇娥李建霖
Owner DONGGUAN HEC MEDICAL INTELLIGENT DEVICE R&D CO LTD
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