Kourilov Vitaly, Steinitz Michael
Experimental Pathology, The Hebrew University-Hadassah Medical School, POB 12272, Jerusalem 91120, Israel.
Anal Biochem. 2002 Dec 15;311(2):166-70. doi: 10.1016/s0003-2697(02)00405-0.
Antigen- and antibody-coated magnetic beads, which are commercially available as activated particles and are readily coated with a variety of molecules, are excellent devices for selecting specific cell populations. We describe an enzyme-linked immunosorbent assay (ELISA)-based method that validates the adsorption of ligand to magnetic beads and verifies the surface accessibility of specific epitopes of the particular ligand. Accordingly, various ligands at a wide range of concentrations were incubated with magnetic beads and adsorption was then assessed using either a specific primary antibody and a secondary alkaline phosphatase-conjugated antibody or a specific primary alkaline phosphatase-conjugated antibody. The method is straightforward and fast and, due to the very low nonspecific background binding, it requires extremely small amounts of beads and ligand. It can be easily performed on newly prepared beads before using them for selection. Magnetic-bead ELISA also confirms the accessibility of specific epitopes on the surface of the beads, which corresponds to the primary antibody used in the assay.
抗原包被磁珠和抗体包被磁珠作为活化颗粒在市场上有售，很容易包被各种分子，是用于选择特定细胞群体的优良工具。我们描述了一种基于酶联免疫吸附测定（ELISA）的方法，该方法可验证配体与磁珠的吸附情况，并验证特定配体特定表位的表面可及性。因此，将各种浓度范围的不同配体与磁珠一起孵育，然后使用特异性一抗和碱性磷酸酶偶联二抗或特异性碱性磷酸酶偶联一抗来评估吸附情况。该方法简单快速，由于非特异性背景结合非常低，因此所需的磁珠和配体量极少。在将新制备的磁珠用于选择之前，可以很容易地对其进行该操作。磁珠ELISA还证实了磁珠表面特定表位的可及性，这与测定中使用的一抗相对应。
