When recombinant proteins go wrong: The hidden pitfall of recombinant protein contamination.

IF 3.7 3区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Cytokine Pub Date : 2024-12-14 DOI:10.1016/j.cyto.2024.156830
Lejla Svraka, Hakim Ben Abdallah, Claus Johansen
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Abstract

Recombinant proteins are critical tools in research; however, their purity is often assumed rather than verified, leading to potential experimental errors. This study aimed to investigate the inflammatory role of recombinant human IL-17F in dermal fibroblasts. Unexpectedly, we discovered with Western blot that recombinant IL-17F from the supplier was contaminated with IL-4, leading to unintended stimulatory effects such as STAT6 phosphorylation and gene induction of CCL26 and IL4R. This contamination led to misinterpretation of data, loss of research time, and erroneous conclusions about IL-17F activity. These findings underscore the critical need for stringent quality control in recombinant protein production and highlight the risks of relying on single-source suppliers. Researchers should remain cautious about potential contamination, ideally validating proteins from multiple suppliers. Our experience illustrates a broader requirement for suppliers to strengthen quality assurance measures, as contaminants can propagate misleading data in the literature and undermine research reproducibility.

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重组蛋白是研究中的重要工具;然而,它们的纯度往往被假定而非验证,从而导致潜在的实验错误。本研究旨在调查重组人 IL-17F 在真皮成纤维细胞中的炎症作用。意想不到的是,我们通过 Western 印迹发现,供应商提供的重组 IL-17F 受到了 IL-4 的污染,导致了意想不到的刺激作用,如 STAT6 磷酸化以及 CCL26 和 IL4R 的基因诱导。这种污染导致了对数据的误读、研究时间的损失以及有关 IL-17F 活性的错误结论。这些发现强调了在重组蛋白生产过程中进行严格质量控制的重要性,并凸显了依赖单一来源供应商的风险。研究人员应该对潜在的污染保持谨慎,最好对来自多个供应商的蛋白质进行验证。我们的经验表明,由于污染物会在文献中传播误导性数据并破坏研究的可重复性,因此更广泛地要求供应商加强质量保证措施。
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来源期刊
Cytokine
Cytokine 医学-免疫学
CiteScore
7.60
自引率
2.60%
发文量
262
审稿时长
48 days
期刊介绍: The journal Cytokine has an open access mirror journal Cytokine: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. * Devoted exclusively to the study of the molecular biology, genetics, biochemistry, immunology, genome-wide association studies, pathobiology, diagnostic and clinical applications of all known interleukins, hematopoietic factors, growth factors, cytotoxins, interferons, new cytokines, and chemokines, Cytokine provides comprehensive coverage of cytokines and their mechanisms of actions, 12 times a year by publishing original high quality refereed scientific papers from prominent investigators in both the academic and industrial sectors. We will publish 3 major types of manuscripts: 1) Original manuscripts describing research results. 2) Basic and clinical reviews describing cytokine actions and regulation. 3) Short commentaries/perspectives on recently published aspects of cytokines, pathogenesis and clinical results.
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