Lagacé J, Arsenault S, Cohen E A
Department of Microbiology and Immunology, Université de Montréal, Quebec, Canada.
J Immunol Methods. 1994 Sep 30;175(1):131-5. doi: 10.1016/0022-1759(94)90339-5.
Short peptides often do not bind well to the plastic surface of microtitre wells in solid-phase immunoassays. To improve the reactivity of synthetic peptides coated on the solid phase, we have developed a highly sensitive, rapid and simple ELISA. This ELISA is based on the treatment of microtitre plates with Alcian blue prepared in acetic acid prior to coating the target peptide. With hyperimmune serum, this treatment increases the specific signal in such a manner that the detection of antibodies needs less antigen than with conventional ELISA. Moreover, the Alcian blue treatment was shown to be particularly effective for the detection of monoclonal antibodies against a synthetic peptide derived from the human immunodeficiency type 1 (HIV-1) vpu protein. These monoclonal antibodies that were not detected in conventional ELISA gave a positive signal up to a 1 in 1000 dilution after Alcian blue treatment of microtitre wells. This assay should be applicable to a variety of synthetic peptides and other types of molecules posing problems in assays requiring their immobilization on solid phases.
在固相免疫测定中，短肽通常与微量滴定板的塑料表面结合不佳。为提高包被在固相上的合成肽的反应性，我们开发了一种高度灵敏、快速且简单的酶联免疫吸附测定（ELISA）。这种ELISA基于在包被目标肽之前，用在乙酸中制备的阿尔辛蓝处理微量滴定板。使用超免疫血清时，这种处理以这样一种方式增加特异性信号，即与传统ELISA相比，检测抗体所需的抗原更少。此外，已证明阿尔辛蓝处理对于检测针对源自人类免疫缺陷病毒1型（HIV-1）vpu蛋白的合成肽的单克隆抗体特别有效。这些在传统ELISA中未检测到的单克隆抗体，在微量滴定孔经阿尔辛蓝处理后，在高达1:1000稀释度时仍给出阳性信号。该测定法应适用于各种合成肽以及在需要将其固定在固相上的测定中存在问题的其他类型分子。
