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血清白蛋白-脂肪酸饱和度试验

Serum albumin-fatty acid saturation test.

作者信息

Gonçalves-de-Albuquerque Cassiano Felippe, Barnese Marcos Roberto Colombo, Soares Mariana Alves, Castro Faria Mauro Velho, Silva Adriana Ribeiro, Castro Faria Neto Hugo Caire, Burth Patrícia, Younes-Ibrahim Mauricio

机构信息

Laboratório de Imunofarmacologia, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro, Brazil.

Laboratório de Imunofarmacologia, Departamento de Bioquímica, Universidade Federal do Estado do Rio de Janeiro, Rio de Janeiro, RJ 20211-010, Brazil.

出版信息

MethodsX. 2019 Aug 24;6:1871-1875. doi: 10.1016/j.mex.2019.08.004. eCollection 2019.

Abstract

Circulating non-esterified fatty acids (NEFA) are toxic to mammalian cells. They increase in diseases such as diabetes and sepsis. Herein we propose a serum albumin-fatty acid saturation test. •We based our test on three methodologies: isoelectric focusing (IF) of human plasma albumin, staining proteins after isoelectric focusing in gels with Coomassie Brilliant Blue, and serum albumin measurement with bromocresol green.•The test consists in the determination of albumin IF and staining with bromocresol green. If albumin is saturated with NEFA, it focuses on lower pH, meaning it is the threshold to bind to them. Excessive NEFA is free and toxic. Many other tests are available for NEFA quantification as NEFA kit assay. All colorimetric assays are used for quantification of NEFA and other tests need expensive equipment to read out the results, and they do not measure albumin levels.•Our method focused on albumin-NEFA saturation instead of just NEFA quantification. Critically ill patients have an alteration in both albumin and NEFA. Therefore, our test undergoes less daytime variation compared to assays that measure absolute NEFA values, allowing a more reliable use as an indicator of albumin-fatty acid saturation and NEFA toxicity.

摘要

循环中的非酯化脂肪酸(NEFA)对哺乳动物细胞有毒性。它们在糖尿病和败血症等疾病中会增加。在此,我们提出一种血清白蛋白-脂肪酸饱和度测试。•我们的测试基于三种方法:人血浆白蛋白的等电聚焦(IF)、用考马斯亮蓝对凝胶中等电聚焦后的蛋白质进行染色以及用溴甲酚绿测定血清白蛋白。•该测试包括测定白蛋白IF并用溴甲酚绿染色。如果白蛋白被NEFA饱和,它会聚焦在较低的pH值,这意味着它是与NEFA结合的阈值。过量的NEFA是游离且有毒的。还有许多其他用于NEFA定量的测试方法可供使用,如NEFA试剂盒检测。所有比色法都用于NEFA的定量,而其他测试需要昂贵的设备来读取结果,并且它们不测量白蛋白水平。•我们的方法侧重于白蛋白-NEFA饱和度,而不仅仅是NEFA定量。重症患者的白蛋白和NEFA都会发生改变。因此,与测量绝对NEFA值的检测方法相比,我们的测试在日间变化较小,作为白蛋白-脂肪酸饱和度和NEFA毒性的指标使用起来更可靠。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d0e/6726920/d478826351c5/fx1.jpg

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