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Preparation method of agarose immune magnetic microspheres and applications thereof

A magnetic microsphere and agarose technology, which is applied in the preparation method of peptide, microsphere preparation, anti-animal/human immunoglobulin, etc., can solve the complex process of monoclonal antibody, highly toxic coupling agent, high cost, etc. problems, achieve the effects of reduced pretreatment time, mild separation and purification conditions, and easy operation

Active Publication Date: 2013-07-24
HARBIN YICAI NEW MATERIAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The present invention aims to solve the technical problems of complex process, low yield, high cost and highly toxic coupling agent of the existing purified monoclonal antibody, thereby providing a preparation method and application of agarose immunomagnetic microspheres

Method used

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  • Preparation method of agarose immune magnetic microspheres and applications thereof
  • Preparation method of agarose immune magnetic microspheres and applications thereof
  • Preparation method of agarose immune magnetic microspheres and applications thereof

Examples

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specific Embodiment approach 1

[0028] Specific embodiment one: the preparation method of a kind of agarose immunomagnetic microsphere of the present embodiment is to carry out according to the following steps:

[0029] 1. Take 10-15g of FeCl3·6H 2 O and 30-50g of sodium oleate are dissolved in a mixed solvent, heated to 60-80°C, and kept for 4-6 hours. After the reaction is completed, take the upper organic solvent and wash it with distilled water three times, and remove the organic solvent by rotary evaporation to obtain ferric oleate. Precursor; wherein, the mixed solvent is formed by mixing ethanol, distilled water and n-hexane in a volume ratio of 60-80:40-60:120-140;

[0030] 2. At room temperature, mix 30-40g of oleic acid, 5-8g of the iron oleate precursor obtained in step 1, and 100-200g of octadecene, and then heat it to 320°C at a rate of 3-5°C per minute. ~340℃, keep warm for 20~50min, then naturally cool to room temperature, add 400~600mL of absolute ethanol, and centrifuge at 3000~5000rpm for ...

specific Embodiment approach 2

[0041] Embodiment 2: This embodiment differs from Embodiment 1 in that: in step 1, heat to 70° C. and keep warm for 4 hours. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0042] Embodiment 3: This embodiment is different from Embodiment 1 or 2 in that: in step 1, the mixed solvent is formed by mixing ethanol, distilled water and n-hexane in a volume ratio of 80:60:140. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a preparation method of agarose immune magnetic microspheres and applications thereof, and relates to a preparation method of microspheres and applications thereof, which solves the technical problems that an existing purified monoclonal antibody is complex in process, is low in yield, and is high in cost, and coupling agents are hypertoxic. The preparation method comprises the following steps of: 1. preparing a ferric oleate precurser; 2. preparing a magnetic fluid; 3. preparing an oil phase and a water phase; 4. preparing agarose magnetic microspheres; 5. preparing activated and crosslinked agarose magnetic microspheres; 6. preparing agarose magnetic microspheres joined with spacer arms; and 7. preparing agarose immune magnetic microspheres. The agarose immune magnetic microspheres provided by the invention are used for purifying genetic engineering recombination trastuzumab monoclonal antibodies. The agarose immune magnetic microspheres prepared by the invention are used for the field of separating and purifying the monoclonal antibodies.

Description

technical field [0001] The invention relates to a preparation method and application of microspheres. Background technique [0002] Monoclonal antibody refers to a homologous antibody produced by a single clone hybridoma cell that only recognizes a specific epitope. It is a special type of antibody prepared artificially. Its essence is globulin, which has various physical and chemical properties, spatial structure and biological activity of globulin. The constant region has a complement binding site, which mediates the function of complement, forms a membrane attack complex, and kills mutated cells. [0003] Monoclonal antibodies have the properties of general antibodies. They are a type of globulin produced when B cells proliferate and differentiate into plasma cells. They exist in body fluids and have immune functions. They can mediate humoral immunity and interact with corresponding antigens (such as pathogens). Specifically bind, and play an immune effect with the part...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/24B01J20/28B01J20/30B01J13/02C07K16/28C07K1/14
Inventor 赵九蓬侯雪梅李垚马丽华潘磊赵长杰
Owner HARBIN YICAI NEW MATERIAL
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