Department of Pathology, The Geisel School of Medicine at Dartmouth, Hanover, NH, United States; Dartmouth-Hitchcock Medical Center, One Medical Center Dr., Lebanon, NH 03756, United States.
Department of Pathology, The Geisel School of Medicine at Dartmouth, Hanover, NH, United States; Dartmouth-Hitchcock Medical Center, One Medical Center Dr., Lebanon, NH 03756, United States.
Clin Chim Acta. 2015 Feb 2;440:169-71. doi: 10.1016/j.cca.2014.11.017. Epub 2014 Nov 21.
Macroenzyme complexes of serum enzymes and antibody can increase the circulating enzymatic activity and may lead to unnecessary additional testing and procedures. Laboratory physicians and scientists need to be aware of techniques to identify macroenzyme complexes when suspected.
To investigate the possibility of a macro-alkaline phosphatase in the serum of a 74 year old male with persistently increased alkaline phosphatase we coupled a protein A/G agarose affinity chromatography technique with isoenzyme electrophoresis to look for the presence of macro-alkaline phosphatase.
The majority of the alkaline phosphatase activity in the patient's serum sample was bound to the column and only a minor fraction (25%) of alkaline phosphatase activity was present in the column flow-through. The alkaline phosphatase activity was also found to co-elute with the immunoglobulins in the patient sample. The alkaline phosphatase activity in a control serum sample concurrently treated in the same manner did not bind to the column and was found in the column flow-through.
The use of protein A/G agarose affinity chromatography is a rapid and simple method that can be applied to the investigation of other macro-enzyme complexes.
血清酶和抗体的宏酶复合物可增加循环酶活性,可能导致不必要的额外检测和操作。实验室医生和科学家需要了解在怀疑存在宏酶复合物时识别它们的技术。
为了研究 74 岁男性血清中是否存在碱性磷酸酶大分子,我们将蛋白 A/G 琼脂糖亲和层析技术与同工酶电泳相结合,以寻找碱性磷酸酶大分子的存在。
患者血清样本中的大部分碱性磷酸酶活性与柱子结合,只有一小部分(25%)的碱性磷酸酶活性存在于柱子的流经液中。在患者样本中,碱性磷酸酶活性也与免疫球蛋白共洗脱。同时以相同方式处理的对照血清样本中的碱性磷酸酶活性未与柱子结合,而是存在于柱子的流经液中。
蛋白 A/G 琼脂糖亲和层析的使用是一种快速简单的方法,可应用于其他宏酶复合物的研究。