Feng Hua-tao, Law Wai-Siang, Yu Li-Jun, Li Sam Fong-Yau
Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore 117543, Republic of Singapore.
J Chromatogr A. 2007 Jul 13;1156(1-2):75-9. doi: 10.1016/j.chroma.2006.12.077. Epub 2006 Dec 23.
The application of quantum dots in capillary electrophoresis immunoassay was studied for the first time. Quantum dots were conjugated with antibody and subsequently tested by electrophoretic separation of free antibody and antibody-antigen complex. Antibody was fluorescently labeled by quantum dots via conjugation procedures and its electrophoretic characteristics were effectively modified due to the attachment of quantum dots. The determination of human IgM by direct CE based immunoassay could be easily achieved by simply changing the pH value of separation buffer. Polymer additive influenced the separation too but the effect was not as significant as buffer pH adjustment. Satisfactory separation of complex from free antibody could be achieved with 20mM sodium tetraborate as separation buffer, at pH 9.8. The immunoassay application of quantum dots in CE offers considerable advantages and can be readily applied to other large bio-molecules.
首次研究了量子点在毛细管电泳免疫分析中的应用。量子点与抗体结合,随后通过游离抗体和抗体 - 抗原复合物的电泳分离进行检测。通过结合程序,量子点对抗体进行荧光标记,并且由于量子点的附着,其电泳特性得到有效改变。通过简单改变分离缓冲液的pH值,基于直接毛细管电泳的免疫分析可以轻松实现人IgM的测定。聚合物添加剂也影响分离,但效果不如缓冲液pH调节显著。以20mM硼酸钠作为分离缓冲液,在pH 9.8时,可以实现复合物与游离抗体的满意分离。量子点在毛细管电泳中的免疫分析应用具有相当大的优势,并且可以很容易地应用于其他大型生物分子。