用于快速筛选杂交瘤培养物中抗雌激素受体抗体产生的荧光酶联免疫吸附测定法。
Fluorometric ELISA method for rapid screening of anti-estrogen receptor antibody production in hybridoma cultures.
作者信息
Wang K C, Leung B S
出版信息
J Immunol Methods. 1985 Nov 28;84(1-2):279-90. doi: 10.1016/0022-1759(85)90435-1.
A micro-fluorometric ELISA which required only 50 microliter of reagent in each step is reported here for rapid detection of anti-estrogen receptor (ER) antibody production by hybridoma cultures. In this study, several factors which influence the sensitivity of ELISA, e.g., incubation time, substrates, microtiter plates, concentration of coating antigens, choice and concentration of specific antibody and enzyme were evaluated. We observed that micro-fluorometric ELISA using 4-methylumbelliferyl phosphate-phosphatase system and MicroFluor W plates was the most sensitive; it detected readily 1 ng coating antigens or 1 ng antibody during a 15 min incubation. This system was used successfully for the detection of anti-ER antibodies in mouse immune sera, and in spent media from in vitro immunized spleen cells or hybridoma cells. This method is simple, sensitive, rapid and reliable for ER or anti-ER antibody determination.
本文报道了一种微量荧光酶联免疫吸附测定法（ELISA），该方法每一步仅需50微升试剂，用于快速检测杂交瘤培养物中抗雌激素受体（ER）抗体的产生。在本研究中，评估了几个影响ELISA灵敏度的因素，如孵育时间、底物、微量滴定板、包被抗原浓度、特异性抗体和酶的选择及浓度。我们观察到，使用4-甲基伞形酮基磷酸-磷酸酶系统和MicroFluor W板的微量荧光ELISA最为灵敏；在15分钟孵育期间，它能轻松检测到1纳克包被抗原或1纳克抗体。该系统已成功用于检测小鼠免疫血清以及体外免疫脾细胞或杂交瘤细胞的培养上清液中的抗ER抗体。该方法对于ER或抗ER抗体的测定简单、灵敏、快速且可靠。
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