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首页> 外文会议>Symposium on Analytical Ultracentrifugation >Combination of preparative and analytical ultracentrifugation methods for the investigation of low-density lipoprotein aliquots to prepare radioactive-labelled lipoproteins
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Combination of preparative and analytical ultracentrifugation methods for the investigation of low-density lipoprotein aliquots to prepare radioactive-labelled lipoproteins

机译：用于研究低密度脂蛋白等分试样的制备和分析超速离心方法以制备放射性标记脂蛋白的研究

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Low-density lipoprotein (LDL) can be radiolabelled with different techniques and various tracers (99mTc, 123I, 125I, 131I, 111In, 67 Ga) and can be used as a tool for the noninvasive exploration of a variety of disorders of lipoprotein metabolism. Density-gradient centrifugation allows the simultaneous isolation of the major lipoprotein density classes, i.e. very low density lipoproteins, LDLs, high-density lipoproteins, forming discrete bands of lipoproteins in preparative tubes. The analysis of the principal lipid constituents was done in isolated fractions. The cholesterol and triglyceride values in the fractions were determined by enzymatic methods. The fraction limits of the bands were defined by means of prestaining the serum lipoproteins. Aliquots of the fractions were taken for Schlieren analysis after adjusting the density and underlayering with a salt solution, in a spinning ultracentrifugation capillary band-forming cell. We obtained quantitative results by measuring the Schlieren areas between the sample curves and the reference baseline curve by computerised numerical and graphical techniques. The decomposition of the integrated curve was carried out using a nonlinear regression program followed by deconvolution algorithm analysis in order to determine the parameters of the constituent Gaussian subclasses. With the LDL particle concentration technique, we measured the concentration of LDL and analysed the polydispersity of the aliquots for the preparing of the radioactive-labelled lipoproteins. LDL is the major transport protein for endogenous cholesterol in human plasma. Radioiodination of LDL was performed using the iodogen method and LDL was radiolabelled with 99mTc using sodium dithionite as a reducing agent. The development of intracellularly trapped ligands permitted the sites of in vivo degradation of LDL to be identified. The methods described proved to be useful for a clear and immediate visual presentation of the concentration values of lipoproteins in the whole spectra and for the identification of the heterogeneity of lipoproteins subfractions. Combination of preparative and analytical ultracentrifugation methods allow lipoprotein aliquots to be investigated before and after radioactive labelling with isotopes to identify labelled materials.
机译：低密度脂蛋白（LDL）可以用不同的技术和各种示踪剂（99mTC，123i，125i，131i，111in，67Ga）放射性标记，并且可以用作非侵入性探索脂蛋白代谢的各种疾病的工具。密度梯度离心允许同时分离主要脂蛋白密度等级，即非常低密度脂蛋白，LDL，高密度脂蛋白，在制备管中形成脂蛋白的离散带。在分离的级分中进行了对本质脂质成分的分析。通过酶法测定级分中的胆固醇和甘油三酯值。带的分数限制通过染色血清脂蛋白来定义。在调节密度和底层的纺丝超离心式毛细管带状带状电池中，在调节密度和底层后，对Schlieren分析进行等分试样。我们通过通过计算机数值和图形技术测量样品曲线和参考基线曲线之间的Schlieren区域获得定量结果。通过非线性回归程序进行集成曲线的分解，然后进行解卷积算法分析，以确定组成高斯子类的参数。利用LDL颗粒浓度技术，我们测量了LDL的浓度，并分析了等分试样的多分散性以制备放射性标记的脂蛋白。 LDL是人血浆内源性胆固醇的主要运输蛋白。使用碘固体方法进行LDL的放射性碘化，使用二硫酸钠作为还原剂，用99MTC放射性标记的LDL。细胞内被捕获的配体的发展允许鉴定LDL的体内降解的位点。所描述的方法证明是有用的，可用于清楚和立即视觉呈现整个光谱中脂蛋白浓度值和鉴定脂蛋白偶联剂的异质性。制备和分析超速离心方法的组合允许在用同位素的放射性标记之前和之后进行脂蛋白等分试样以鉴定标记的材料。

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