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开发一种敏感的侧流免疫试验,使用复合子单链可变片段与酸纤维素结合蛋白融合

Ngoc Minh Nguyen1, Ryosuke Mukai1, Makoto Nita1

  • 1Faculty of Molecular Chemistry and Engineering, Kyoto Institute of Technology, Kyoto 606-8585, Japan.

Analytical chemistry
|December 15, 2025
PubMed

Insights

我们开发了一个新的横流免疫测试 (LFIA) 平台,使用与结合蛋白融合的复合抗体片段来提高诊断性能. 这一创新提高了抗原检测的灵敏度和可复制性在 point-of-care测试.

科学领域:

  • 生物技术是生物技术.
  • 免疫技术是一种免疫技术.
  • 材料科学 材料科学 材料科学

背景情况:

  • 侧流免疫检测 (LFIA) 对于快速的护理点诊断至关重要.
  • 传统的抗体固定在酸纤维素 (NC) 膜上具有局限性.
  • 为了提高LFIA的业绩,需要加强固定化策略.

研究的目的:

  • 开发一个创新的LFIA平台,使用与NC结合蛋白 (NBPs) 基因融合的抗体片段.
  • 为了增强抗体固定稳定性和NC膜上的定向.
  • 提高LFIAs的灵敏度和可重复性.

主要方法:

  • 对21个候选NBP进行查,确定乳酸铁素 (LF) 是优质的.
  • 单链可变片段-LF (scFv-LF) 融合蛋白的构造和表达.
  • 使用补充性确定区域 (CDR) 插入scFv-LF变体.
  • 在点滴,ELISA和侧流量测试中评估性能.

主要成果:

  • 乳二显示出强大而稳定的吸附到NC膜上.
  • scFv-LF融合蛋白在广泛的pH范围内显示出高的抗原结合活性.
  • CDR移植的scFv-LF变体 (例如B1R/C2R) 显著增强了抗原活性和信号强度.
  • B1R/C2R scFv-LF 融合实现了流感B核体蛋白的100倍较低的检测极限.

结论:

  • 该scFv-LF融合平台能够在NC膜上实现稳定和定向的抗体固定.
  • 这种方法大大提高了LFIA的敏感性和可重复性.
  • 模块化的CDR-grafting策略为下一代诊断提供了快速定制的便利.
抽象的

No abstract available in PubMed .

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