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当前位置: 首页 > 文章 > 不同磁珠对过敏原蛋白纯化效果的比较研究 山东农业大学学报(自然科学版) 2017,48 (6) 853-857
Position: Home > Articles > Comparison of Different Magnetic Beads for Purification of Allergen Protein Journal of Shandong Agricultural University(Natural Science Edition) 2017,48 (6) 853-857

不同磁珠对过敏原蛋白纯化效果的比较研究

作  者：
李文娇;阎俊;杜娟;黄天娇;卢瑛
关键词：
磁珠;过敏原纯化;原肌球蛋白;免疫磁分离技术
摘  要：
本文以南美白对虾的主要过敏原为纯化对象,选用2种商业磁珠构建免疫磁珠,利用免疫磁分离技术纯化过敏原,采用SDS-PAGE、Western blot、蛋白定量对纯化产物进行鉴定,探讨了不同免疫磁珠对过敏原的纯化效果.结果表明,磁珠A的抗体偶联量明显高于磁珠B.此外,免疫磁珠A对抗原的结合率和洗脱率分别为免疫磁珠B的1.5、1.8倍;免疫磁珠A对过敏原捕获率为47.9%,而免疫磁珠B仅为17.6%.对比两种磁珠的理化及其结构特点,发现粒径大、分散性差的磁珠在偶联过程会发生聚集,降低抗体偶联效率,从而影响纯化效果.结果显示,分散性好、抗体偶联率高的磁珠更适宜蛋白质的分离纯化.本研究为以后过敏原的纯化提供了一定的理论基础与实验依据.
译  名：
Comparison of Different Magnetic Beads for Purification of Allergen Protein
关键词：
Magnetic bead%allergen purification%tropomyosin%immunomagnetic separation
摘  要：
In this study, the main allergen tropomyosin (Tm) of Penaeus vannamei was used as the purification object. Two commercial magnetic nanoparticles were selected to build immunomagnetic beads. The allergen was purified by immunomagnetic separation technology. SDS-PAGE, western blot and protein quantification methods were employed to identify the purified sample , and the effect of different immune magnetic beads for the purification of allergen was compared. Results showed that antibody coupling amount of magnetic bead A was higher than that of magnetic beads B. Besides, the combination and elution rates of immune magnetic beads A for antigen was 1.5 and 1.8 times than that of immune magnetic bead B, respectively;the capture rate of allergen by immunomagnetic beads A was 47.9%, while immunomagnetic beads B was only 17.6%. By comparing the physicochemical and structural properties of the two beads, the magnetic bead with larger bead-size and poor dispersion was gathered during the coupling process, which reduced the antibody coupling efficiency and affect the purification effect. This paper showed that magnetic bead with good dispersion and high antibody coupling rate is more suitable for the separation and purification of protein. This study provides a theoretical basis and experimental basis for the purification of allergens in the future.

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