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Magnetic bead method for quickly purifying antibody

A magnetic bead method, magnetic bead technology, applied in organic chemistry, peptide preparation methods, chemical instruments and methods, etc.

Inactive Publication Date: 2013-09-04
HENAN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Commonly used methods for antibody purification include caprylic ammonium sulfate precipitation, ion exchange chromatography and SPA affinity chromatography. A large number of chemical reagents and experimental instruments (such as centrifuges, purification chromatography workstations, etc.), preparation of various buffer solutions, and multi-step operations (such as centrifugation, precipitation, chromatography purification, etc.) are required. The whole process takes a long time and is effective. During working hours, the method of human operation or machine purification, the number of purified samples is limited

Method used

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  • Magnetic bead method for quickly purifying antibody
  • Magnetic bead method for quickly purifying antibody
  • Magnetic bead method for quickly purifying antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Purification of Antibodies from Human Whole Blood and Pig Serum by Protein A Magnetic Beads

[0037] 1. Antibody purification

[0038] (1) Take 0.5mL of human whole blood (or 0.1mL of pig serum) into a 2mL EP tube;

[0039] (2) Dilute human whole blood 1-3 times with pH 7.4 PBS buffer;

[0040] (3) Add 50uL Protein A magnetic beads (vortex or pipette to mix the magnetic beads before use);

[0041] (4) Vortex for 10s, incubate at room temperature for 10min (during this period, vortex twice);

[0042] (5) Place the centrifuge tube on the magnetic stand, suck off the supernatant after 1 min (the centrifuge tube is kept on the magnetic stand), and be careful not to attract the magnetic beads;

[0043] (6) Add 500uL pH 7.4 PBS buffer, vortex for 5s, and wash the ProteinA magnetic beads;

[0044] (7) Place the centrifuge tube on the magnetic stand, suck off the supernatant after 1 min (the centrifuge tube is kept on the magnetic stand), and be careful not to ...

Embodiment 2

[0065] Example 2: Purification of antibodies from pig serum (1 mL) by Protein A magnetic beads—consecutive use for multiple times

[0066] 1. Antibody purification

[0067] (1) Take 0.2mL of pig serum into a 2mL EP tube;

[0068] (2) Dilute 1-3 times with pH 7.4 PBS buffer;

[0069] (3) Add 50uL ProteinA magnetic beads (vortex or use a pipette to mix the magnetic beads before use);

[0070] (4) Vortex for 10s, incubate at room temperature for 10min (during this period, vortex twice);

[0071] (5) Place the centrifuge tube on the magnetic stand, suck off the supernatant after 1 min (the centrifuge tube is kept on the magnetic stand), and be careful not to attract the magnetic beads;

[0072] (6) Add 500uL pH 7.4 PBS buffer, vortex for 5s, and wash the ProteinA magnetic beads;

[0073] (7) Place the centrifuge tube on the magnetic stand, suck off the supernatant after 1 min (the centrifuge tube is kept on the magnetic stand), and be careful not to attract the magnetic bead...

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Abstract

The invention relates to the biological and pharmaceutical industry, and discloses a magnetic bead method for quickly purifying an antibody. The method is characterized in that by using an affinity chromatography principle, carboxyl magnetic beads are activated, and then connected with stphylococcl protein A (SPA) capable of being combined with Fc (fragment crystalline) of IgG (intravenous gamma globulin) molecules of human and various mammals; and obtained Protein A magnetic beads can be used for purifying the antibody in serum, ascites or a cell culture fluid. The Protein A magnetic beads are high in specificity, quick and convenient in the aspect of antibody purification, and can be widely applied to the biological and pharmaceutical industry.

Description

technical field [0001] The invention relates to biological and pharmaceutical industries, and can more conveniently and quickly purify IgG class antibodies from animal ascites, serum or cell culture. Background technique [0002] Commonly used methods for antibody purification include caprylic ammonium sulfate precipitation, ion exchange chromatography, and SPA affinity chromatography. These methods require a large number of chemical reagents and experimental instruments (such as centrifuges, purification chromatography, etc.) in the process of antibody purification. Workstation, etc.), prepare a variety of buffers, and perform multi-step operations (such as centrifugation, precipitation, chromatographic purification, etc.), the whole process takes a long time, and within the effective working time, the method of human operation or machine purification, its purification Sample size is limited. Compared with the traditional purification method, the magnetic bead method has o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00C07K1/14
Inventor 李华梁永波陆启玉
Owner HENAN UNIVERSITY OF TECHNOLOGY
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