Application of lectin-magnetic carrier coupling compound in separation of glycosylated exosome
A magnetic carrier and lectin technology, applied in the direction of carrier binding/immobilization of peptides, solid adsorbent liquid separation, separation methods, etc., can solve the problems of large steric hindrance, rupture of exosome vesicles, and low separation efficiency, and achieve The effect of reducing steric hindrance, reducing steric hindrance, and improving separation efficiency
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Embodiment example 1
[0054] Example 1 Preparation of lectin-magnetic carrier conjugated complex
[0055] The lectin-magnetic carrier conjugated complex includes: a magnetic carrier; and a lectin coupled to the outside of the magnetic carrier, which is prepared by coupling the lectin and the magnetic carrier under specific conditions, and is mainly used for separating clinical samples The glycosylated exosomes obtained after isolation are complete in morphology. in:
[0056] Lectins are mainly used: pineapple agglutinin (Jacalin), peanut agglutinin (PNA), pea agglutinin (VVA and / or VVL), concanavalina agglutinin (ConA), lentil agglutinin (LCA), wheat germin (WGA), soybean lectin (SBA), kidney bean lectin (PVL), snail lectin (HAA and / or HPA), or any combination of two or more lectins, the main reasons are: The combination of different lectins can realize the separation of various types of glycosylated exosomes, such as: LCA, AAL for the separation of fucosylated exosomes; ConA, PVL, SBA, WGA, AAL ...
Embodiment example 2
[0070] Example 2 Preparation of Glycosylated Exosome Separation Composition
[0071] The present invention provides a composition for separating glycosylated exosomes, which specifically includes: a lectin-magnetic carrier conjugated complex, which is used for washing and removing non-specifically bound and unglycosylated exosomes during the separation process. and other impurities, and / or eluate for elution of glycosylated exosomes specifically bound to lectin-magnetic carrier conjugated complexes, individually packaged and present in a set or in the kit.
[0072] In this example, the above-mentioned lectin-magnetic carrier conjugated complex is the LCA-agarose magnetic bead solution with a volume ratio of 50% obtained in Example 1.
[0073] The above cleaning solution is a metal-salt ion-free cleaning buffer or purified water, and can optionally be a metal-salt-free cleaning buffer, such as: the main component includes 10-200mM metal-salt-free TRIS-HCl buffer with a pH of 7...
Embodiment example 3
[0075] Example 3 Method of using glycosylated exosome isolation composition
[0076] The present invention also provides a method for separating glycosylated exosomes, which mainly includes the main experimental steps of using the composition for separating glycosylated exosomes in Example 2. Please refer to the schematic diagram of the separation principle for details. figure 1 "Schematic diagram of the separation of glycosylated exosomes by lectin-magnetic carrier conjugated complexes".
[0077] The detailed experimental steps of the present invention include:
[0078] 1. Preparation before the experiment
[0079] Self-provided equipment or equipment: magnetic stand for manual separation of glycosylated exosomes; or use the fully automatic agarose magnetic bead separation instrument of the group company and its subsidiaries, mainly for automatic glycosylated exosome separation separation, to achieve the purpose of saving manpower. The automatic separation of glycosylated ...
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