Improved methods for magnetic purification of malaria parasites and haemozoin
Improved methods for magnetic purification of malaria parasites and haemozoin
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DOI：
10.1186/1475-2875-9-17
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发表时间：
2010-01-14
期刊：
影响因子：
3
通讯作者：
DeRisi, Joseph L.
中科院分区：
文献类型：
--
作者：
Kim, Charles C.;Wilson, Emily B.;DeRisi, Joseph L.
背景：疟原虫在红细胞内生长周期中，在宿主血红蛋白的催化下产生游离血红素。为了使三价铁的氧化毒性最小化，游离血红素分子聚合成生物矿物β-血红素（通常称为血红素）。疟原虫色素晶体是顺磁性的，并且这种性质可以用于晚期寄生虫的纯化，因为它们含有比早期寄生虫和未感染细胞更大的疟原虫色素晶体。最初开发用于抗体介导的细胞纯化目的的市售磁体广泛用于此目的。由于这些方法不一定是最佳的寄生虫纯化，磁场强度和寄生虫纯化过程中的产量和质量的关系进行了探索。方法：廉价的稀土钕磁铁与商业上可获得的一次性柱，以探讨磁场强度和回收率之间的关系游离的疟原虫色素和感染的红细胞（iRBC）。游离色素的产率随着磁场强度的增加而几乎线性地增加，直到测试的最强磁场（8，500高斯）。更强的磁场也改善了iRBC的回收，对寄生虫活力没有不利影响。一个内部建造的磁性支架，为75美元的材料，产生了上级的结果相比，更昂贵的商业products.Conclusions：现有的协议的磁性纯化游离的疟原虫色素和iRBC的结果在次优的产量。廉价的高强度钕磁铁提供了一个更好的选择，导致更高的产量，对寄生虫的生存能力没有不利影响。
Background: Malaria parasites generate free haem upon catabolism of host haemoglobin during their intraerythrocytic growth cycle. In order to minimize oxidative toxicity of the ferric iron, the free haem molecules are polymerized into the biomineral beta-haematin (commonly referred to as haemozoin). Haemozoin crystals are paramagnetic, and this property can be exploited for the purification of late stage parasites as they contain larger haemozoin crystals than early stage parasites and uninfected cells. Commercially available magnets that were originally developed for the purpose of antibody-mediated cell purification are widely used for this purpose. As these methods are not necessarily optimized for parasite purification, the relationship between magnetic field strength and the quantity and quality of yield during parasite purification was explored.Methods: Inexpensive rare-earth neodymium magnets with commercially available disposable columns were employed to explore the relationship between magnetic field strength and recovery of free haemozoin and infected erythrocytes (iRBCs).Results: Yields of free haemozoin increased nearly linearly with increasing magnetic field strength to the strongest fields tested (8,500 Gauss). Stronger magnetic fields also improved the recovery of iRBCs with no detrimental effects on parasite viability. An in-house constructed magnetic stand, built for $75 in materials, produced superior results when compared with much more expensive commercial products.Conclusions: Existing protocols for the magnetic purification of free haemozoin and iRBCs result in sub-optimal yields. Inexpensive high-strength neodymium magnets offer a better option, resulting in higher yields with no detrimental effects on parasite viability.