Sankiewicz Anna, Laudanski Piotr, Romanowicz Lech, Hermanowicz Adam, Roszkowska-Jakimiec Wiesława, Debek Wojciech, Gorodkiewicz Ewa
Department of Electrochemistry, Institute of Chemistry, University of Bialystok, 15-443 Bialystok, Poland.
Department of Perinatology, Medical University of Bialystok, 15-276 Bialystok, Poland.
Anal Biochem. 2015 Jan 15;469:4-11. doi: 10.1016/j.ab.2014.09.021. Epub 2014 Oct 13.
We have developed a new method for highly selective determination of the ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) concentration using a surface plasmon resonance imaging (SPRI) technique and two different biosensors. UCH-L1 was captured from a solution by immobilized specific rabbit monoclonal antibody or specific LDN-57444 inhibitor due to formation of receptor-UCH-L1 complex on the biosensor surface. The analytically useful dynamic response range of both biosensors is between 0.1 and 2.5ng/ml. The detection limit is 0.06ng/ml for the biosensor with antibody and 0.08ng/ml for the biosensor with inhibitor. Biosensors based on both antibody and inhibitor were found to be suitable for quantitative determination of the UCH-L1 and exhibit good tolerance to the potential interferents. Both biosensors gave comparable results in the range of 0 to 0.20ng/ml for plasma samples and 0.30 to 0.49ng/ml for cerebrospinal fluid samples. To validate the new methods, comparative determination of UCH-L1 by the commercial enzyme-linked immunosorbent assay (ELISA) kit was performed. In general, in terms of UCH-L1 concentration, a good correlation between SPRI and ELISA was found. The developed biosensors can be used successfully for the determination of UCH-L1 in body fluids.
我们开发了一种新方法,可使用表面等离子体共振成像(SPRI)技术和两种不同的生物传感器来高度选择性地测定泛素羧基末端水解酶L1(UCH-L1)的浓度。由于在生物传感器表面形成受体-UCH-L1复合物,UCH-L1通过固定化的特异性兔单克隆抗体或特异性LDN-57444抑制剂从溶液中捕获。两种生物传感器的分析有用动态响应范围均在0.1至2.5ng/ml之间。抗体生物传感器的检测限为0.06ng/ml,抑制剂生物传感器的检测限为0.08ng/ml。基于抗体和抑制剂的生物传感器均适用于UCH-L1的定量测定,并且对潜在干扰物具有良好的耐受性。对于血浆样品,两种生物传感器在0至0.20ng/ml范围内以及对于脑脊液样品在0.30至0.49ng/ml范围内均给出了可比的结果。为了验证新方法,使用商业酶联免疫吸附测定(ELISA)试剂盒对UCH-L1进行了比较测定。总体而言,就UCH-L1浓度而言,发现SPRI和ELISA之间具有良好的相关性。所开发的生物传感器可成功用于测定体液中的UCH-L1。