Pillai M R, John C S, Troutner D E
Department of Chemistry, University of Missouri, Columbia 65211.
Bioconjug Chem. 1990 May-Jun;1(3):191-7. doi: 10.1021/bc00003a003.
We report the labeling of human gamma globulin with the 105Rh complex of a new pentadentate bifunctional ligand, 1,7-bis(2-hydroxybenzyl)-4-(p-aminobenzyl)diethylenetriamine. Complexes of this ligand with 105Rh were prepared by refluxing rhodium carrier spiked with 105Rh at pH9 in bicarbonate buffer. The complex was treated with an excess concentration of thiophosgene to prepare the isothiocyanate derivative which was extracted into CHCl3. The CHCl3 extract was dried and dissolved in DMF and reacted with a borate solution of human gamma globulin. Labeling yields were generally high and varied from 73% to 93%, depending upon the concentration of human gamma globulin and the isothiocyanate derivative of the complex used. The overall recovery of rhodium activity varied from 59% to 75% without taking into account activity lost due to decay. The conjugation reaction was complete by 4 h. From 0.4 to 8.5 atoms of Rh could be incorporated per molecule of protein by this method. The activated isothiocyanate complex did not show any degradation when stored at room temperature for up to 4 days and then used for conjugation.
我们报道了用一种新型五齿双功能配体1,7-双(2-羟基苄基)-4-(对氨基苄基)二乙烯三胺的¹⁰⁵Rh络合物标记人γ球蛋白的方法。该配体与¹⁰⁵Rh的络合物是通过在pH9的碳酸氢盐缓冲液中回流加入¹⁰⁵Rh的铑载体来制备的。用过量的硫光气处理该络合物以制备异硫氰酸酯衍生物，然后将其萃取到CHCl₃中。将CHCl₃萃取液干燥并溶解于DMF中，再与人类γ球蛋白的硼酸盐溶液反应。标记产率通常较高，根据所用人类γ球蛋白的浓度和络合物的异硫氰酸酯衍生物不同，产率在73%至93%之间变化。在不考虑因衰变而损失的活性的情况下，铑活性的总回收率在59%至75%之间。共轭反应在4小时内完成。通过这种方法，每分子蛋白质可掺入0.4至8.5个Rh原子。活化的异硫氰酸酯络合物在室温下储存长达4天然后用于共轭反应时，未显示出任何降解。
