Ji Tao, Muenker M Catherine, Papineni Rao V L, Harder John W, Vizard Douglas L, McLaughlin William E
Carestream Molecular Imaging, Carestream Health, Inc., New Haven, Connecticut 06511.
Bioconjug Chem. 2010 Mar 17;21(3):427-35. doi: 10.1021/bc900295v. Epub 2010 Feb 17.
IgG antibodies were conjugated to Kodak X-Sight nanospheres to develop fluorescent-labeled antibodies using two different synthetic routes: one involving the DTT reduction method, and the other involving Traut's Reagent modification method. These two methods result in different conjugation efficiencies and different performances in antigen detection. Western blotting shows that the nanosphere-IgG antibody conjugates synthesized using the DTT reduction method are more immunospecific than the conjugates synthesized using Traut's Reagent modification method. In addition, the conjugates synthesized using DTT reduction also show higher antigen detection sensitivity than other commercially available fluorescent-IgG antibody conjugates, including Alexa Fluor, Qdot, and CyDye conjugates.
将IgG抗体与柯达X-Sight纳米球偶联,通过两种不同的合成途径开发荧光标记抗体:一种涉及二硫苏糖醇(DTT)还原法,另一种涉及特劳特试剂修饰法。这两种方法在抗原检测中产生不同的偶联效率和不同的性能。蛋白质印迹法表明,使用DTT还原法合成的纳米球-IgG抗体偶联物比使用特劳特试剂修饰法合成的偶联物具有更高的免疫特异性。此外,使用DTT还原法合成的偶联物在抗原检测灵敏度方面也高于其他市售荧光-IgG抗体偶联物,包括Alexa Fluor、量子点和CyDye偶联物。