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Micro-fluidic chip based on magnetic bead coated antibody and method for capturing cardiac markers

A microfluidic chip, myocardial marker technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of reconstituted antigen capture, detachment from coating substrates, etc.

Active Publication Date: 2016-03-23
北京乐普诊断科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to achieve the purpose of instant detection, the immune substrate is fully mixed through high mass transfer in the microchannel, and then there are problems such as redissolving the coated antibody in the fluid system and being captured by the antigen in the fluid and detached from the coated substrate.

Method used

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  • Micro-fluidic chip based on magnetic bead coated antibody and method for capturing cardiac markers
  • Micro-fluidic chip based on magnetic bead coated antibody and method for capturing cardiac markers
  • Micro-fluidic chip based on magnetic bead coated antibody and method for capturing cardiac markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0129] In this example, cardiac troponin I (cTnI) will be described.

[0130] 1) Making immunomagnetic beads coated with antibodies, the making of immunomagnetic beads coated with antibodies that can specifically bind to cTnI protein in blood includes:

[0131] Immunomagnetic beads (Cat.: LSKMAGA10, Merck Millipore) were equilibrated with MES buffer (2-(N-morpholine)ethanesulfonic acid) until the pH of the solution was 4-5 (preferably 4.5) and the ionic strength was 0.1M. Add EDC and NHS to the balanced solution respectively until the concentrations of EDC and NHS in the solution are both 1 mg / mL, react for 20 minutes and then centrifuge to discard the supernatant;

[0132] Reconstitute the precipitate after discarding the supernatant with the MES buffer (before equilibration), and add antibody 8E10 (Cat.: 4T13, Hytest) until the concentration of 8E10 is 0.5 mg / mL;

[0133] Mix and coat for 1 hour, then add BSA (Cat.: 10735108001, Roche) to a BSA concentration of 1% to termin...

Embodiment 2

[0145] In this embodiment, myoglobin MYO is taken as an example for illustration.

[0146] 1) The production of immunomagnetic beads coated with antibodies that can specifically bind to the MYO protein in blood includes:

[0147] The immunomagnetic beads are equilibrated with PB (phosphate buffer solution) buffer until the pH of the solution is 7.4 (7.0-8.0 is acceptable) and the ionic strength is 0.1M.

[0148] Add EDC and NHS to the balanced solution respectively until the concentration of both EDC and NHS in the solution is 1 mg / mL. After reacting for 20 minutes, centrifuge and discard the supernatant.

[0149] Reconstitute the precipitate after discarding the supernatant with the PB buffer (before equilibration), add antibody (Cat.: 4M23, Hytest) to the reconstituted solution to a 4E2 concentration of 0.5mg / mL, mix and coat After 1 h, BSA (bovine serum albumin) was added until the final concentration of BSA was 1%, and the reaction was terminated.

[0150] 2) Labeled wit...

Embodiment 3

[0161] This example illustrates C-reactive protein.

[0162] 1) The preparation of immunomagnetic beads coated with antibodies that can specifically bind to the CRP protein in blood includes: the immunomagnetic beads are equilibrated with MES buffer until the pH of the solution is 4.5-5.5 (preferably 5), and the ionic strength is 0.1M;

[0163] Then add EDC and NHS respectively, until the final concentration of both EDC and NHS is 1 mg / mL, and react for 20 minutes. Discard the supernatant by centrifugation, and redissolve the precipitate after discarding the supernatant with the PBS buffer (before equilibration), add antibody CRP135 (Cat.: 4C28, Hytest) to a final concentration of CRP135 of 0.5mg / mL, mix After 1 h, BSA was added until the final concentration of BSA was 1%, and the reaction was terminated.

[0164] 2) Colloidal gold (self-made) labeling for CRP antigen labeling marker: 40nm colloidal gold solution with K 2 CO 3 The solution was equilibrated to a pH of 9 (all...

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Abstract

The embodiment of the invention discloses a micro-fluidic chip based on a magnetic bead coated antibody and a method for capturing cardiac markers. The micro-fluidic chip based on the magnetic bead coated antibody and the method for capturing cardiac markers are applied to the field of immunomagnetic bead coated binding antibodies, and prevent coated antibodies from being redissolved in a fluid system or being captured by antigens in fluid and disengaged from a base material. The micro-fluidic chip comprises a sample injection chamber, a Y-shaped structure channel and recycling chambers. The Y-shaped structure channel comprises a funnel-shaped channel at the upper portion and a columnar channel at the lower portion. An immunolabelling module is arranged at the bottom of the funnel-shaped channel. A blood filtering module is arranged at the upper portion of the immunolabelling module. The bottom end of the columnar channel at the lower portion is forked to form a pair of parallel immunity channels. The tail end of each immunity channel is connected with one recycling chamber. The columnar channel is provided with an upper cavity and a lower cavity. The upper cavity is a strong magnetism cavity, and the lower cavity is a quality control cavity.

Description

technical field [0001] The invention relates to the field of immunomagnetic beads coated with binding antibodies, in particular to a microfluidic chip based on magnetic beads coated with antibodies and a method for capturing myocardial markers. Background technique [0002] At present, microfluidic technology has made important progress in the fields of biochemical analysis, immunodiagnosis, and environmental detection. For example, under the premise of finely controlling the flow rate, the antigen-antibody reaction can be controlled to achieve high-sensitivity control of the immune response. The myocardial antibody is coated in the reaction microchamber by means of van der Waals force, electrostatic adsorption, polymer embedding, etc., for antigen capture. In order to achieve the purpose of instant detection, the immune substrate is fully mixed through high mass transfer in the microchannel, and then there are problems such as redissolving the coated antibody in the fluid s...

Claims

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Application Information

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IPC IPC(8): G01N33/533
CPCG01N33/533
Inventor 胡飞熊晶张单单邱笑违余占江
Owner 北京乐普诊断科技股份有限公司
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