在通过AlphaFold,HADDOCK和MD模拟验证的ScFv-CD99结合亲和度中,BioPhi驱动的人性优化的可塑性
Kanokporn Sornsuwan1,2, Thanathat Pamonsupornwichit2, On-Anong Juntit1,2
1Office of Research Administration, Chiang Mai University, Chiang Mai 50200, Thailand.
Computational and structural biotechnology journal
|February 3, 2025
概括
对抗CD99单链可变片段 (ScFvs) 的BioPhi导向人性化改善了结构稳定性,但显示出不同的结合亲和性. 尽管增强了人性,但所有变体都带来了高的免疫性风险,这强调了治疗抗体开发中需要保持平衡的需要.
科学领域:
- 生物技术是生物技术.
- 免疫学 免疫学 免疫学
- 结构生物学 结构生物学
背景情况:
- 抗体碎片的人性化对于减少治疗应用中的免疫性至关重要.
- 针对CD99的单链可变片段 (ScFvs) 正在探索治疗潜力.
- 在保持结合亲和力和稳定性的同时,优化 ScFv 人性是关键的挑战.
研究的目的:
- 通过使用BioPhi引导的人性化来增强抗CD99 ScFv的人性.
- 评估人化对抗CD99 ScFvs.的结构稳定性,结合亲和力和免疫性的影响.
- 调查人性,结构完整性和工程 ScFvs. 的功能性能之间的相互作用.
主要方法:
- 生物Phi引导的人化,以创建HuScFvMT99/3BP和HuScFvMT99/3HY的变种.
- 使用AlphaFold2,AlphaFold3和HADDOCK进行分子动力学模拟,以评估结合的自由能量 (ΔGbinding) 和结构稳定性 (RMSD).
- 生物层干涉计,ELISA和流动细胞计,以实验验证结合亲和和和相互作用. 用于免疫性风险评估的PITHA分析.
主要成果:
- 与野生类型 (WT) 相比,HuScFvMT99/3BP和HuScFvMT99/3HY的结构稳定性得到改善.
- 结合亲和结果在模拟方法中不一致;生物层干涉测量表明WT具有最高亲和力 (KD = 1.35 × 10−7 M),其次是BP和HY变体.
- 所有变种都表现出高免疫性风险,尽管HuScFvMT99/3HY在其互补性决定区域 (CDR) 中表现出更好的人性和特定的结构特征.
结论:
- 生物Phi引导的人性化可以增强ScFv的人性和结构稳定性,但可能会损害结合亲和力.
- 为了获得最佳的治疗抗体候选人,需要平衡人类的功能有效性和最小化免疫性.
- 需要进一步的优化策略来开发安全有效的人性化抗CD99 ScFv治疗方法.
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