偏好的子抗体对NOTCH3免疫原体C末端的反应
Soo Jung Lee1,2, Mitchell B Gasche1, Connor J Burrows1
1Department of Neurology, University of Michigan, Ann Arbor, MI, 48109, USA.
Scientific reports
|June 6, 2023
概括
子抗体通常针对的C端,限制它们对全蛋白的使用. 这项研究发现,超过70%的NOTCH3抗体更喜欢C端表位,阻碍了更广泛的应用.
科学领域:
- 免疫学 免疫学 免疫学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 从免疫接种产生的抗体被广泛用于生物研究.
- 由于抗体反应偏差,针对特定蛋白质存在挑战.
- 之前在小鼠中的观察表明,在幽默反应中偏爱C端向.
研究的目的:
- 为了调查偏好的子抗体对免疫原的C-终端反应的频率.
- 评估子产生的抗体对人类NOTCH3的有用性,用于生物研究.
主要方法:
- 对人类的10个序列产生23个多克隆抗体 NOTCH3.3.
- 使用具有C端延伸或截断的重组NOTCH3序列来表征抗体活性.
- 在免疫细胞化学应用中对抗体性能的评估.
主要成果:
- 在生成的多克隆抗体中,超过70% (16分之 23) 显示出C端偏好.
- 这些抗体主要向免疫性的自由卡尔基.
- 抗体对缺乏自由C端或在它之前截断的重组蛋白质的反应性降低或不存在.
- 免疫细胞化学证实了对表达自由C端的细胞倾向的反应性.
结论:
- 子表现出显著的倾向产生抗体对NOTCH3衍生的C端表位.
- 这种C端偏差可以限制这些抗体对本地NOTCH3蛋白的有效性.
- 讨论了克服这种偏差的潜在策略,以提高抗体生成效率.
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