Exosomes
have become the most ideal analysis target for liquid
biopsy since they carry a large amount of genetic materials. The study
on exosomes has great significance for cancer diagnosis and prognosis.
However, the extremely low concentration renders the development of
a robust exosomes enrichment technique, with the merits of low nonspecific
cell adhesion, high-capture efficiency, and easy nondestructive release
of captured exosomes, of vital significance. We successfully designed
and developed a novel Tim4@ILI-01 immunoaffinity flake material. First,
a strongly hydrophilic ILI-01 MOFs matrix material was fabricated
with cationic ionic liquid 1,3-bis(4-carboxybutyl)imidazolium bromide
as the organic ligand. The nonspecific adsorption of the ILI-01 MOFs
material was only 0.7% after two washings with a neutral buffer. Moreover,
based on the inherent abundant carboxyl groups on the ILI-01 MOFs
flake, they can be facilely functionalized with an anti-Tim4 antibody
with the bonding efficiency of 82.4%. The capture efficiency of the
developed Tim4@ILI-01 immunoaffinity material for exosomes reached
85.2%, which is 5.2 times higher than that via the gold standard ultracentrifugation
method. Furthermore, based on the Ca2+-dependent characteristic
of the binding between the Tim4@ILI-01 immunoaffinity material and
phosphatidylserine (PS) on the surfaces of exosomes, the captured
exosomes can be easily released with the addition of a chelating agent
under neutral eluent conditions. Thus, the captured exosomes maintained
good biological activity. The developed Tim4@ILI-01 immunoaffinity
flake was successfully applied for enrichment of exosomes from serums
of healthy persons and lung adenocarcinoma patients. The levels of
the expressed CD44 gene significantly changed under different stages
of lung adenocarcinoma cancer. All these results demonstrate that
the Tim4@ILI-01 immunoaffinity flake is a robust enrichment material
and has a good potential in practical clinical applications.