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PMID: 23025585 Published · ppublish English Journal Article

Noble polymeric surface conjugated with zwitterionic moieties and antibodies for the isolation of exosomes from human serum.

Bioconjugate chemistry ·Vol. 23 ·No. 10 ·2012-10-17 ·Pages 2114-20

Kim G, Yoo CE, Kim M, Kang HJ, Park D, Lee M, Huh N

Abstract

New zwitterionic polymer-coated immunoaffinity beads were developed to resist nonspecific protein adsorption from undiluted human serum for diagnostic applications of exosomes. A zwitterionic sulfobetaine monomer with an amine functional group was employed for simple surface chemistry and antifouling properties. An exosomal biomarker protein, epithelial cell adhesion molecule (EpCAM), was selected as a target molecule in this work. The beads were coated with polyacrylic acids (PAA) for increasing biorecognition sites, and protein G was then conjugated with carboxylic acid groups on the surfaces for controlling EpCAM antibody orientation. The remaining free carboxylic acid groups were modified with sulfobetaine moieties, and anti-EpCAM antibody was finally introduced. The amount of anti-EpCAM on the beads was increased by 40% when compared with PAA-uncoated beads. The surfaces of the beads exhibited near-net-zero charge, and nonspecific protein adsorption was effectively suppressed by sulfobetaine moieties. EpCAM was captured from undiluted human serum with almost the same degree of efficiency as from PBS buffer solution using the newly developed immunoaffinity beads.

MeSH Terms
Acrylic Resins/chemistry Adsorption Antibodies/chemistry,immunology Antigens, Neoplasm/immunology Cell Adhesion Molecules/immunology Cell Fractionation/methods Epithelial Cell Adhesion Molecule Exosomes/metabolism Humans Magnets/chemistry Microspheres Surface Properties
Chemicals
Acrylic Resins Antibodies Antigens, Neoplasm Cell Adhesion Molecules Epithelial Cell Adhesion Molecule carbopol 940
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Kim Gahee
Bio Research Center, Samsung Advanced Institute of Technology (SAIT), Mt. 14-1, Nongseo-dong, Yongin-si, 446-712, Gyeonggi-do, South Korea.
Yoo Chang Eun
Kim Myoungsoon
Kang Hyun Ju
Park Donghyun
Lee Myoyong
Huh Nam
Article Info
Journal
Bioconjugate chemistry
Abbr.
Bioconjug Chem
ISSN
1520-4812
Published
2012-10-17
Epub
2012-00-08
Pages
2114-20
Language
English
Region
United States
NLM ID
9010319
Subset
IM
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