Laboratory of Research in Biotechnology and Hemoderivatives, Department of Pharmaceutical Sciences, Federal University of Pernambuco, 50670-420 Recife, Pernambuco, Brazil.
Laboratory of Research in Biotechnology and Hemoderivatives, Department of Pharmaceutical Sciences, Federal University of Pernambuco, 50670-420 Recife, Pernambuco, Brazil; Laboratory of Bioactive Technology, Department of Morphology and Animal Physiology, Federal Rural University of Pernambuco, 52171-900 Recife, Pernambuco, Brazil.
Int J Biol Macromol. 2021 Jul 1;182:2056-2065. doi: 10.1016/j.ijbiomac.2021.05.179. Epub 2021 Jun 2.
Precipitation of blood products from plasma fractionation has played a fundamental role in the industrial purification of important therapeutic products. Only a few studies have been reported by using tannins as proteins precipitant agent from whole plasma while, several conditions have been analyzed. Here, we decided to verify the effect of the temperature on the precipitation process of plasma proteins using tannic acid (TA). Plasma proteins were precipitated with tannic acid by using different temperature incubations. Subsequently, the protein-TA complex was analyzed by SDS-PAGE and quantified. In addition, the protein activity of the complex was measured after heating, as well as the structural changes of the complexes were accompanied by thermogravimetric analysis, differential scanning calorimetry and circular dichroism. In all conditions tested, tannic acid was able to precipitate without selectively separating the proteins in the mixture by using different temperatures during the precipitation process. Furthermore, the protein concentration from the plasma precipitate was not affected by different temperatures and the plasma precipitate was able to dissolve fibrin clots in vitro.
从血浆分离物中沉淀血液制品在重要治疗产品的工业纯化中起着至关重要的作用。虽然已经有一些使用单宁作为蛋白质沉淀剂从全血浆中沉淀的研究报告,但只有少数条件得到了分析。在这里,我们决定使用单宁酸(TA)来验证温度对血浆蛋白沉淀过程的影响。使用不同的温度孵育,用单宁酸沉淀血浆蛋白。随后,通过 SDS-PAGE 分析和定量分析蛋白-TA 复合物。此外,在加热后测量复合物的蛋白活性,以及通过热重分析、差示扫描量热法和圆二色性伴随结构变化。在所有测试的条件下,单宁酸能够沉淀,而无需通过沉淀过程中使用不同的温度选择性分离混合物中的蛋白质。此外,不同温度对血浆沉淀中的蛋白质浓度没有影响,并且血浆沉淀能够在体外溶解纤维蛋白凝块。