In various embodiments methods are provided for identifying and/or quantifying one or more antigens of interest (biomarkers) on the surface of cell- or tissue-specific exosomes. In an illustrative embodiments the methods comprise: i) incubating a population of exosomes with one or more tissue-specific antibodies that bind an antigen specific to a tissue or cell type of interest that produces exosomes, where the tissue specific antibodies are attached to acceptor bead or magnetic beads so the antibodies bind exosomes displaying the antigen; ii) obtaining a purified population of exosomes bound by the tissue specific antibodies with and/or without photocleavable linker based technology; iii) incubating a test subset of the isolated tissue-specific exosomes with acceptor beads attached to test antibodies that bind an antigen of interest thereby binding exosomes that display the antigen of interest and a control subset with negative control acceptor beads; v) incubating the test subset of isolated exosomes and the control subset of exosomes with a donor-bearing antibody that binds an exosome specific antigen; and vi) detecting a signal produced upon illumination of the control subset and/or the test; and vii) detecting the antigen(s) of interest.
在各种实施例中,提供了用于识别和/或定量细胞或组织特异性外泌体表面上的一个或多个感兴趣的抗原(
生物标志物)的方法。在一个说明性实施例中,该方法包括:i)将外泌体群体与一个或多个组织特异性
抗体孵育,这些
抗体与特定于产生外泌体的感兴趣组织或细胞类型的抗原结合,其中这些组织特异性
抗体附着在受体珠或磁珠上,使
抗体结合显示抗原的外泌体;ii)获得由组织特异性
抗体结合的外泌体的纯化群体,有时使用光解链技术;iii)将分离的组织特异性外泌体的测试子群体与附着有结合感兴趣抗原的测试
抗体的受体珠孵育,从而结合显示感兴趣抗原的外泌体,以及一个带有阴性对照受体珠的对照子群体;v)将分离的外泌体的测试子群体和对照子群体与结合外泌体特异性抗原的供体
抗体孵育;vi)在照射对照子群体和/或测试子群体时检测产生的信号;以及vii)检测感兴趣的抗原。