Stability of important antibodies for kidney disease: pre-analytic methodological considerations.
Stability of important antibodies for kidney disease: pre-analytic methodological considerations.
复制标题
肾脏疾病重要抗体的稳定性：分析前方法学考虑
作者：
Han Q;Li S;Fu B;Liu D;Wu M;Yang X;Cai G;Liu Z;Chen X;Zhu H
近年来，人们认识到循环抗体作为肾脏疾病生物标志物的重要性。然而，没有研究系统地描述了将抗体作为肾脏疾病的生物标志物的样品制备和储存方法。目前尚不清楚反复的冻融循环、物理干扰、不同温度或不同时间的储存、溶血或混浊的血清样本是否会影响抗体的测量。本研究的目的是探讨在各种相关的临床和研究条件下血清样本中与肾脏疾病相关的抗体的稳定性。方法将血清样品在以下不同条件下保存：重复冻融循环（1、6或12次）、长期保存（- 80℃保存7或12个月）、物理干扰（1或8小时）、4℃保存（1、3或6周）和室温保存（1或7天）。采用酶联免疫吸附试验（ELISA）评价抗磷脂酶A2受体（anti-PLA2R）、抗肾小球基底膜、抗髓过氧化物酶和抗蛋白酶3抗体的稳定性。结果反复冻融循环对血清样品中上述抗体的稳定性无显著影响。溶血和浑浊血清的ELISA读数分别有升高和降低的趋势。在- 80°C的长期保存和物理干扰对密封血清样品中抗pla2r抗体的稳定性都没有显著影响。在4℃下保存6周以上或在室温下保存7天以上的未密封血清样品中，大多数抗体浓度升高。我们的研究结果表明，上述用于肾脏疾病生物标志物的循环抗体具有稳定的物理化学性质、结构和免疫反应性，因此它们不受重复冻融循环、物理干扰或在- 80°C下长期储存的影响。然而，对于溶血、浑浊和未密封的血清样本，ELISA读数倾向于改变。
Background The importance of circulating antibodies as biomarkers of kidney disease has recently been recognized. However, no study has systematically described the methodology of sample preparation and storage regarding antibodies as biomarkers of kidney disease. It remains unknown whether repetitive freeze-thaw cycles, physical disturbances, storage at different temperatures or for different periods of time, or haemolytic or turbid serum samples affect antibody measurements. The aim of this study was to investigate the stabilities of antibodies associated with kidney disease in serum samples under various relevant clinical and research conditions. Methods We stored serum samples in the following different conditions: repetitive freeze-thaw cycles (1, 6 or 12 times), long-term storage (7 or 12 months at −80 °C), physical disturbance (1 or 8 h), and storage at 4 °C (1, 3 or 6 weeks) and room temperature (1 or 7 days). The stabilities of the anti-phospholipase A2 receptor (anti-PLA2R), anti-glomerular basement membrane, anti-myeloperoxidase and anti-proteinase 3 antibodies were evaluated with enzyme-linked immunosorbent assays (ELISA). Results We found that repetitive freeze-thaw cycles did not have a significant effect on the stabilities of the abovementioned antibodies in clear serum samples. The ELISA readings of haemolytic and turbid serum samples tended to increase and decrease, respectively. Neither long-term storage at −80 °C nor physical disturbance had a significant effect on anti-PLA2R antibody stability in sealed serum samples. The concentrations of most of these antibodies increased in unsealed serum samples that were stored at 4 °C for more than 6 weeks or at room temperature for more than 7 days. Discussion Our findings revealed that the abovementioned circulating antibodies that are used as biomarkers for kidney disease had stable physicochemical properties, structures and immunoreactivities such that they were not influenced by repetitive freeze-thaw cycles, physical disturbances or long-term storage at −80 °C. However, the ELISA readings tended to change for haemolytic, turbid and unsealed serum samples.
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影响因子：
13.6
作者：
Yang, Rui;Hellmark, Thomas;Wang, Hai-yan
通讯作者：
Wang, Hai-yan
影响因子：
3.8
作者：
Hodgkinson, Verity S.;Egger, Sam;Sitas, Freddy
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影响因子：
1.8
作者：
Michaut, Lydia;Laurent, Nathalie;Deckert-Salva, Fabienne
通讯作者：
Deckert-Salva, Fabienne
影响因子：
2.6
作者：
Gomez-Morales, Maria Angeles;Ludovisi, Alessandra;Pozio, Edoardo
通讯作者：
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影响因子：
5
作者：
WATANABE, F;TSUBOTA, N;KAWASHIMA, H
通讯作者：
KAWASHIMA, H