Biguzzi S
Boll Ist Sieroter Milan. 1977 Nov 30;56(5):489-94.
A method for the improvement of commercial antisera that consist of ammonium sulfate precipitated gamma-globulins labelled with fluorescein isothiocyanate, is presented. By a simple desalting on Sephadex G25 followed by a DEAE-cellulose stepwise chromatography, it is easy to recover only the fluoresceinated immunoglobulins of optimal fluorescein/protein ratio. The aspecificities due to the other fluoresceinated (mainly albumin) and also to the hypo- and hyper-conjugated gamma-globulins are then avoided. Even if eventual defects as regards the intrinsic characteristics of the antisera are not modified by this process, the improvement obtained is stringent and particularly evident in the indirect immunofluorescent staining of tissue sections. In consideration of the excellent quantitative recovery, this simple procedure is recommended to people that use commercial reagents for studies that are beyond the qualitative limits of routine diagnostic immunofluorescent tests.
本文介绍了一种改进商业抗血清的方法，该抗血清由硫酸铵沉淀的异硫氰酸荧光素标记的γ-球蛋白组成。通过在葡聚糖凝胶G25上进行简单的脱盐，然后进行DEAE-纤维素分步色谱，很容易仅回收具有最佳荧光素/蛋白质比率的荧光免疫球蛋白。这样就避免了由于其他荧光标记物（主要是白蛋白）以及低共轭和高共轭γ-球蛋白引起的非特异性。即使该过程不会改变抗血清固有特性方面的最终缺陷，但所获得的改进是显著的，并且在组织切片的间接免疫荧光染色中尤为明显。考虑到出色的定量回收率，对于那些使用商业试剂进行超出常规诊断免疫荧光测试定性范围研究的人，推荐使用这个简单的程序。
