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The Preparation of Immunomagnetic Microspheres and Application in Breast Cancer Diagnose Research

Author: HanZhiGang
Tutor: HouZhanMing；QiuGuangLiang
School: Inner Mongolia Normal
Course: Botany
Keywords: Immunomagnetic microspheres Avidin Circulating tumor cells Cell separation
CLC: R737.9
Type: Master's thesis
Year: 2010
Downloads: 100
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Abstract

Cancer is currently the several major diseases plaguing human life, and into increasingly hyperplasia situation. The average annual incidence of cancer in about 22%. Therefore, the treatment of cancer is a the people pressing task. Most of the cancers detected in the current medical facilities are all advanced discovered relatively late basic-proliferation of cancer cells at this time, the drug is difficult to treat. The most important thing is to fight cancer, early detection, the most early cancer can be cured. First disorderly growth of cancerous tumor cells to lose the ability of mutual adhesion loose tumor cells can enter the blood and lymph vessels, into the blood circulation or lymphatic circulation to achieve the transfer of cells called circulating tumor cells (circulating through the blood circulation tumor cells; CTCs). Tumor early transfer of circulating tumor cells (CTCs) can be planted in distant organs along the circulatory system, and gradually formed metastases. Using existing conventional empirical diagnostic tool but can not \Therefore, research and development of a new method to detect metastatic tumor cells is very necessary. Many studies have proved that the application detected a small amount of blood circulating tumor cell clusters (CTCs) to analyze the results, specific prognosis on for each speculated independent patients, to assess the accuracy and effectiveness of treatment response can be significantly improved. Obviously, the level of individual cells of the peripheral blood blood circulating tumor cells (CTCs) more sensitive than imaging analysis. Recently developed technology combined with the \Breast cancer cells are found in human breast relative to other cancer cells in vivo more vulnerable to external mechanical forces and thus easier to fall off the formation of circulating tumor cells. Further specific immune magnetic microspheres to capture circulating tumor cells of early breast cancer early detection relative to the early detection of other cancers are more convenient and accurate. Immunomagnetic microspheres micron This article will focus on the synthesis and preparation of immunomagnetic beads, focusing on the expedition preparation conditions, the removal efficiency of the hybrid cell, the capture rate of the circulating tumor cells. First, the particle size of the magnetic microspheres, magnetic response surface carboxyl content analysis to filter out more suitable for the preparation of immunomagnetic microspheres beads; then synthesized by covalent coupling method immunomagnetic microsphere complexes; longer This composite body to capture the target cell, to achieve heteroaryl cell removal and the enrichment of the circulating tumor cells. Specific research methods and conclusions are summarized as follows: using transmission electron microscopy, scanning electron microscopy, electron microscopy determination of the particle size of the microspheres, to observe the surface morphology. Characterization of magnetic microspheres (Magnetic polymer microspheres) by measuring the conductivity of surface carboxyl content. Pick out the particle size is evenly distributed around 10μm, better into a ball, the surface carboxyl group content of 0.1 to 0.5 mmol / g of magnetic microspheres. In certain oscillation conditions, by covalent coupling SYSTEM prepared immune magnetic microspheres composite body, spectrophotometer, the rate of determination of the binding of avidin (neutravidin). Experimental results show that the reaction temperature in the avidin - magnetic microsphere preparation process at 4 ℃, avidin concentration 0.2mg/ml reaction in pH 5.8 PBS reaction environment 2h, the avidin load 25 to 55mg / g. Preparation of biotinylated antibody the BNHS and antibody mass ratio 1:7 when combined with the highest rate of the coupling of the highest rates of avidin and biotin molar ratio of 1:3. Anti-CD45 immunomagnetic microspheres leukocyte removal rate can be as high as 78%, the the magnetic microspheres recovery rate of 98%. Therefore, the specific immune magnetic microspheres capture the blood circulating tumor cell technology has good application prospects in the early diagnosis of breast cancer.

The Preparation of Immunomagnetic Microspheres and Application in Breast Cancer Diagnose Research

Abstract

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