Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 1208, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jchromb.2022.123408
Keywords
Polyhedral oligomeric silsesquioxane; Graphene; HCIC ligand; Click reaction; Antibody
Funding
- National Natural Science Foundation of China [21874106, 22174108, 21605122]
- Natural Science Foundation of Shaanxi Province [2021JM-310]
- Open Foundation of Key Laboratory of Synthetic and Natural Functional Molecule of the Ministry of Education [338050067]
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In this study, a novel graphene-based composite was prepared and successfully applied for the separation of antibodies in serum. The composite exhibited high selectivity towards antibodies and effectively hindered the non-specific adsorption of other proteins.
In this work, we prepared a novel graphene-based composite by combination of mercapto-end capped polyhedral oligomeric silsesquioxane (POSS-SH) with graphene oxide (GO) through covalent bonding process. The composite was further conjugated with 4-vinyl pyridine (4-VP) through thiol-ene click reaction to fabricate pseudomercaptoethyl pyridine functionalized POSS-graphene composite (G-POSS-S-VP). The physicochemical properties were confirmed via FT-IR, TGA, XPS, and TEM characterizations and a high ligand density of ca. 87.3 mu mol m- 2 was obtained. G-POSS-S-VP composite displayed high selectivity toward antibody, i.e., immunoglobulin G (IgG) in this case based on the hydrophobic charge induced chromatography (HCIC) mechanism. A high adsorption efficiency of ca. 90% and large adsorption capacity of 500 mg g-1 were achieved for IgG at pH 7.4. At the same time, the non-specific adsorptions of co-existing proteins on graphene substrate were obviously hindered. The practical applicability of G-POSS-S-VP composite as adsorbent was verified by selective separation of IgG from human serum. SDS-PAGE assay and MALDI-TOF-MS analysis clearly demonstrated the successful isolation of IgG with high purity.
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