Engineered recombinant single-chain fragment variable antibody for immunosensors
Engineered recombinant single-chain fragment variable antibody for immunosensors
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DOI：
10.1021/ac0507690
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发表时间：
2005-11-01
影响因子：
7.4
通讯作者：
Zeng, XQ
中科院分区：
文献类型：
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作者：
Shen, ZH;Mernaugh, RL;Zeng, XQ
设计了一种重组单链可变区(ScFv)抗体(命名为A10B)，该抗体在连接ScFv重链和轻链的连接肽中含有两个组氨酸。成功研制了一种基于单链抗体的压电免疫传感器。A10B单链抗体结合到金压电免疫传感器表面的定位正确，保持了抗原结合活性，并在较高的表面浓度下偶联。这些结果以及早期使用含有半胱氨酸的单链抗体的研究结果表明，连接氨基酸在接头序列上的位置对单链抗体具有很好的耐受性，并且不干扰单链抗体的抗原结合活性。该传感器的灵敏度为2.3+/-0.15 nm(n=4)，线性范围为2.3×10(-9)~3.3×10(-8)M。该传感器还用于研究抗原/单链抗体结合的动力学和热力学。
A recombinant single-chain fragment variable (scFv) antibody (designated A10B) was engineered to contain two histidines within the linker peptide used to join the scFv heavy and light chains. A piezoimmunosensor using the scFv was successfully developed. A10B scFv bound to the gold piezoimmunosensor surface were correctly oriented, retained antigen-binding activity, and coupled at high surface concentration. These results, and results obtained from an earlier study using an scFv containing a linker cysteine, suggest that the location on the linker sequence in which the amino acids were incorporated was well tolerated by the scFv and did not interfere with scFv antigen-binding activity. The scFv-modified QCM sensor was thoroughly characterized and used to specifically detect antigen in crude serum sample and had a sensitivity of 2.3 +/- 0.15 nM (n = 4) with a linear range over 2.3 x 10(-9)-3.3 x 10(-8) M. The piezoimmunosensor was also used to study the kinetics and thermodynamics of antigen/scFv antibody binding.