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Successful Protein Extraction from Over-Fixed and Long-Term Stored Formalin-Fixed Tissues

机译:从过固和长期保存的福尔马林固定组织中成功提取蛋白质

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摘要

One of the major breakthroughs in molecular pathology during the last decade was the successful extraction of full-length proteins from formalin-fixed and paraffin-embedded (FFPE) clinical tissues. However, only limited data are available for the protein extraction efficiency of over-fixed tissues and FFPE blocks that had been stored for more than 15 years in pathology archives. In this study we evaluated the protein extraction efficiency of FFPE tissues which had been formalin-fixed for up to 144 hours and tissue blocks that were stored for 20 years, comparing an established and a new commercial buffer system. Although there is a decrease in protein yield with increasing fixation time, the new buffer system allows a protein recovery of 66% from 144 hours fixed tissues compared to tissues that were fixed for 6 hours. Using the established extraction procedure, less than 50% protein recovery was seen. Similarly, the protein extraction efficiency decreases with longer storage times of the paraffin blocks. Comparing the two buffer systems, we found that 50% more proteins can be extracted from FFPE blocks that were stored for 20 years when the new buffer system is used. Taken together, our data show that the new buffer system is superior compared to the established one. Because tissue fixation times vary in the routine clinical setting and pathology archives contain billions of FFPE tissues blocks, our data are highly relevant for research, diagnosis, and treatment of disease.
机译:在过去的十年中,分子病理学的重大突破之一是从福尔马林固定和石蜡包埋(FFPE)的临床组织中成功提取了全长蛋白质。但是,对于在病理档案中存储了15年以上的过度固定组织和FFPE块的蛋白质提取效率,只有有限的数据可用。在这项研究中,我们比较了已建立的和新型的商业缓冲系统,对经过福尔马林固定长达144小时的FFPE组织和已储存20年的组织块的蛋白质提取效率进行了评估。尽管随着固定时间的增加,蛋白质产量会降低,但与固定6小时的组织相比,新的缓冲系统可以使144小时固定的组织中的蛋白质回收率达到66%。使用已建立的提取程序,发现蛋白质回收率不到50%。同样,随着石蜡块保存时间的延长,蛋白质的提取效率也会降低。比较这两种缓冲液系统,我们发现使用新的缓冲液系统时,可以从存储了20年的FFPE块中提取出50%的蛋白质。综上所述,我们的数据表明,新的缓冲系统比已建立的系统优越。由于常规临床设置中的组织固定时间各不相同,并且病理学档案中包含数十亿个FFPE组织块,因此我们的数据与疾病的研究,诊断和治疗高度相关。

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