Quantitative and qualitative analysis of stability for 16 serum immunoregulators over 50 freeze–thaw cycles

IF 1.6 4区医学 Q1 ANTHROPOLOGY

American Journal of Human Biology Pub Date : 2024-04-29 DOI:10.1002/ajhb.24087

R. N. Chowdhury, A. Armato, E. Culver, L. Shteynman, C. Kurien, B. Cradin, F. Margolin, T. Nguyen, C. Cardona, N. Kabir, R. M. Garruto, J. K. Lum, K. Wander

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Abstract

Objectives

To evaluate the reliability of data from the assay of bio-archived specimens, a 50-freeze–thaw-cycle (FTC) degradation study of fresh sera was conducted to test the stability of 16 immunoregulators.

Methods

Twenty de-identified serum specimens were obtained from volunteers at United Health Services-Wilson Memorial Hospital. Specimens were stored at −20°C and underwent daily 1 h thawing and subsequent freezing for each FTC over 50 consecutive days. Immunoregulator concentrations were assessed via enzyme-linked immunosorbent assay (ELISA) in participant samples at 2 FTC (baseline), 25 FTC, and 50 FTC. Specific immunoregulators observed in the study were C-reactive protein (CRP), interleukin (IL)-1α, 4, 6, 8, 10, monocyte chemoattractant protein-1 (MCP-1, CCL2), monocyte chemoattractant protein-2 (MCP-2, CCL8), eotaxin-1, thymus-and-activation-regulated chemokine (TARC, CCL17), regulated on activation normal T-cell expressed and secreted (RANTES, CCL5), growth-regulated oncogene-alpha (GRO-α, CXCL1), small inducible cytokine A1 (I-309, CCL1), interferon-gamma (IFN-γ), interferon-gamma inducible protein-10 (IP-10, CXCL10), and tumor necrosis factor-alpha (TNF-α).

Results

Quantitative stability of serum immunoregulators: Serum CRP, IL-8, IL-10, IFN-γ, IP-10, and eotaxin-1 levels appear to be statistically equivalent from baseline to 50 FTC (p ≤ .05). Retention of patterns in serum immunoregulators: patterns across FTC were retained for TARC (age) and CRP, IFN-γ, and MCP-2 (sex).

Conclusions

While the effect of multiple FTC on serum immunoregulator levels may not replicate prolonged freezer storage, the results of this study provide valuable information on the robustness of immunoregulators for research using bio-archived sera.

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对 16 种血清免疫调节剂在 50 次冻融循环中的稳定性进行定量和定性分析

为了评估生物萃取标本检测数据的可靠性，我们对新鲜血清进行了 50 次冷冻-解冻循环（FTC）降解研究，以检测 16 种免疫调节剂的稳定性。标本储存在零下 20 摄氏度，每天解冻 1 小时，然后在连续 50 天内冷冻每种 FTC。在 2 FTC（基线）、25 FTC 和 50 FTC 时，通过酶联免疫吸附试验 (ELISA) 对参与者样本中的免疫调节剂浓度进行评估。研究中观察到的特定免疫调节剂包括 C 反应蛋白 (CRP)、白细胞介素 (IL)-1α、4、6、8、10、单核细胞趋化蛋白-1 (MCP-1，CCL2)、单核细胞趋化蛋白-2 (MCP-2，CCL8)、共济失调素-1、胸腺和激活调节趋化因子 (TARC，CCL17)、RANTES，CCL5）、生长调节癌基因-α（GRO-α，CXCL1）、小诱导细胞因子 A1（I-309，CCL1）、干扰素-γ（IFN-γ）、干扰素-γ诱导蛋白-10（IP-10，CXCL10）和肿瘤坏死因子-α（TNF-α）。结果血清免疫调节因子的定量稳定性：血清 CRP、IL-8、IL-10、IFN-γ、IP-10 和 eotaxin-1 水平从基线到 50 FTC 在统计学上似乎是等同的（p ≤ .05）。血清免疫调节剂模式的保留：TARC（年龄）和 CRP、IFN-γ 和 MCP-2（性别）的模式在不同的 FTC 中保留不变。

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来源期刊

American Journal of Human Biology 生物-生物学

CiteScore

4.80

自引率

13.80%

发文量

124

审稿时长

4-8 weeks

期刊介绍： The American Journal of Human Biology is the Official Journal of the Human Biology Association. The American Journal of Human Biology is a bimonthly, peer-reviewed, internationally circulated journal that publishes reports of original research, theoretical articles and timely reviews, and brief communications in the interdisciplinary field of human biology. As the official journal of the Human Biology Association, the Journal also publishes abstracts of research presented at its annual scientific meeting and book reviews relevant to the field. The Journal seeks scholarly manuscripts that address all aspects of human biology, health, and disease, particularly those that stress comparative, developmental, ecological, or evolutionary perspectives. The transdisciplinary areas covered in the Journal include, but are not limited to, epidemiology, genetic variation, population biology and demography, physiology, anatomy, nutrition, growth and aging, physical performance, physical activity and fitness, ecology, and evolution, along with their interactions. The Journal publishes basic, applied, and methodologically oriented research from all areas, including measurement, analytical techniques and strategies, and computer applications in human biology. Like many other biologically oriented disciplines, the field of human biology has undergone considerable growth and diversification in recent years, and the expansion of the aims and scope of the Journal is a reflection of this growth and membership diversification. The Journal is committed to prompt review, and priority publication is given to manuscripts with novel or timely findings, and to manuscripts of unusual interest.