Lectin-macromolecule carrier coupling compound for separating glycosylated exosome in clinical sample

Abstract

The present invention relates to a lectin-macromolecule carrier coupling compound for separating a glycosylated exosome in a clinical sample. The lectin-macromolecule carrier coupling compound comprises a macromolecule carrier; and lectin coupled to the outer side of the macromolecular carrier. The lectin-macromolecular carrier coupling compound provided by the invention can be used for simply, conveniently, rapidly and accurately separating the glycosylated exosomes in the clinical sample; the compound is simple in operation, high in separation efficiency and good in repeatability, and the separated exosome is complete in form and free of breakage or fragmentation and can be directly used for glycosylated exosome liquid detection or directly used for immunological related detection or directly used for gene detection and analysis by extracting related nucleic acid in the exosome.

Description

technical field [0001] The invention belongs to the technical fields related to biomedicine and bioseparation, and relates to a lectin-macromolecular carrier coupling complex used for separating glycosylated exosomes in clinical samples. Background technique [0002] Exosomes are biologically active tiny vesicles released when cell multivesicular endosomes fuse with the plasma membrane, with a diameter of about 30-150 nm, and are one of the important mediators of intercellular communication; under healthy human physiological conditions Exosomes can transport DNA, protein, mRNA, miRNA and other biologically active substances between cells, participate in various processes such as cell communication, cell migration, tumor cell growth, etc., and complete the transmission of intercellular substances and information. Literature research has found that almost all types of cells secrete exosomes, and exosomes can be extracted from body fluids such as blood, saliva, urine, cerebrosp...

AI Technical Summary

Problems solved by technology

Among them: ultracentrifugation or gradient density centrifugation is based on the small difference in the density of exosomes and other biological components to separate, this method requires special equipment, the demand for samples is relatively large, it takes a long time, and because the characteristics of different clinical samples are different , the stability of the separation effect using this method is poor, and further ultracentrifugation may easily cause vesicle damage and affect subsequent experiments; the principle of the immunomagnetic bead method is that specific proteins on the surface of exosomes, such as CD9 / CD63 / CD81, etc. The immunomagnetic beads with corresponding antibodies are combined for magnetic separation. However, due to the small size of the immunomagnetic beads, the particle size is at the nanometer level and less than or equal to the diameter of the exosomes, so the space for the combination of the immunomagnetic beads and the exosomes is Larger resistance, insufficient binding of exosomes, and low separation efficiency

In addition, the exosomes obtained by immunomagnetic bead separation are eluted with acidic eluent, resulting in incomplete morphology of exosomes.

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