Immunomagnetic separation of human myeloperoxidase using an antibody-mimicking peptide identified by phage display

Title
Immunomagnetic separation of human myeloperoxidase using an antibody-mimicking peptide identified by phage display
Author
이은규
Keywords
Human myeloperoxidase; Phage display; Antibody-mimicking peptide; Binding affinity; Immuno-binding; Magnetic particle
Issue Date
2017-09
Publisher
ELSEVIER SCIENCE BV
Citation
JOURNAL OF BIOTECHNOLOGY, v. 257, Page. 118-121
Abstract
Phage display biopanning is a powerful in vitro selection process for screening and identifying peptides that bind to a target protein of interest. With the aim of replacing antibodies in immuno-diagnostic applications, we identified peptides whose binding characteristics mimicked those of anti-human myeloperoxidase (hMPO), a biomarker for acute cardiac diseases. Based on ELISA results from four phage clones, we selected and chemically synthesized a 12-mer peptide (SYIEPPERHRHR). Quartz crystal microbalance and surface plasmon resonance analyses revealed that the molar binding equilibrium ratio of the synthesized peptide was 0.023, approximately 43-fold lower than that of the anti-hMPO antibody. The dissociation constant (K-d) was 57 nM, which was comparable to that of the native antibody (83 nM). Next, we biotinylated the peptide at its N-terminus and attached the biotinylated peptide to the surface of streptavidin-coated magnetic particles to assess its ability to selectively capture hMPO. The binding equilibrium data were similar to the previous analyses; specifically, around 0.021 mol peptide bound to 1 mol of hMPO. Antigen capture was found to be selective and to be relatively little influenced by the presence of human serum albumin (HSA), an abundant constituent of serum. Our work demonstrates the potential of immunomagnetic isolation to achieve selective capture of a low-concentration antigen from complex solutions such as serum. (C) 2016 Elsevier B.V. All rights reserved.
URI
https://www.sciencedirect.com/science/article/pii/S0168165616316443http://repository.hanyang.ac.kr/handle/20.500.11754/72364
ISSN
0168-1656; 1873-4863
DOI
10.1016/j.jbiotec.2016.12.010
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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