Cyclic peptide ligand with high binding capacity for affinitypurification of immunoglobulin G = Cyclic peptide ligand를이용한 IgG 정제

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Title
Cyclic peptide ligand with high binding capacity for affinitypurification of immunoglobulin G = Cyclic peptide ligand를이용한 IgG 정제
Author(s)
H J Kang; W Choe; Jeong Ki Min; Young Mi Lee; B M Kim; S J Chung
Bibliographic Citation
Journal of Chromatography A, vol. 1466, pp. 105-112
Publication Year
2016
Abstract
The rapidly increasing implementation of antibodies in therapeutic and diagnostic applications has neces-sitated the development of antibody production and purification technologies for both academic andindustrial usage. Bacterial Protein A and Protein G are known to bind antibodies with high affinity andhave facilitated the isolation and purification thereof. Recently, small peptide ligands (i.e. IgG Fc domain-binding peptides, FcBP) that specifically bind to the Fc-domain of antibodies were reported. In the presentstudy we describe the development of a reusable high affinity column for antibody purification utilizingimmobilized FcBP, comprising 13 amino acids residues, on a sepharose resin. In addition to FcBP, Cys toSer substituted FcBP (FcBP-Ser), reduced FcBP (FcBP-Red), commercial Protein A and Protein G resins,packed into columns, were evaluated for antibody purification. All these columns except the FcBP-Serone showed good binding capacity for a humanized IgG (trastuzumab) and a chimeric IgG (cetuximab).The column packed with FcBP-Red allowed antibody purification at a less acidic pH (pH 4.8) than wasrequired for the other ligand affinity columns used in our experiments (i.e., pH 3.2 for Protein G and FcBPcolumns, and pH 3.5 for Protein A column, respectively). Utilizing the FcBP column, antibodies from swinehuman sera were isolated with a purity of 95%. Interestingly, the FcBP column could be easily regeneratedand operated without loss of efficiency for up to 60 runs, the maximum number of runs performed in thepresent study.
Keyword
Antibody purificationAffinity columnIgG Fc domain-binding peptide (FcBP)Protein AProtein G
ISSN
0021-9673
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.chroma.2016.09.007
Type
Article
Appears in Collections:
Division of Biomedical Research > Biotherapeutics Translational Research Center > 1. Journal Articles
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