Adamczyk M, Gebler J C, Mattingly P G
Abbott Laboratories, Diagnostics Division, Abbott Park, Illinois 60064, USA.
Bioconjug Chem. 1996 Jul-Aug;7(4):475-81. doi: 10.1021/bc960035h.
Fifteen hapten-bovine serum albumin (BSA) conjugates were prepared from five commercially available activated haptens. Each hapten was coupled to BSA at three different ratios. The conjugates were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two mass spectrometry (MS) methods: matrix-assisted laser desorption ionization (MALDI) and liquid chromatography-electrospray ionization (LC-ESI). SDS-PAGE was useful in detecting protein cross-linking, but not assessing hapten density. MALDI-MS and LC-ESI-MS gave comparable qualitative results, but LC-ESI-MS provided a clearer representation of the distribution of hapten-protein species present in the conjugates. Conjugate species substitute with up to 25 haptens per BSA were recorded by LC-ESI-MS.
由五种市售活性半抗原制备了十五种半抗原 - 牛血清白蛋白（BSA）缀合物。每种半抗原以三种不同比例与BSA偶联。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS - PAGE）和两种质谱（MS）方法对缀合物进行表征：基质辅助激光解吸电离（MALDI）和液相色谱 - 电喷雾电离（LC - ESI）。SDS - PAGE可用于检测蛋白质交联，但不能评估半抗原密度。MALDI - MS和LC - ESI - MS给出了可比的定性结果，但LC - ESI - MS能更清晰地呈现缀合物中存在的半抗原 - 蛋白质种类的分布。通过LC - ESI - MS记录了每个BSA最多可替代25个半抗原的缀合物种类。
