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相关概念视频

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or...
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Development of a Lateral Flow Immunochromatographic Strip for Rapid and Quantitative Detection of Small Molecule Compounds
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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
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概括

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.

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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.