저널명 : Journal of Biotechnology, 289, 88-92 (2019) 
저자 : Soi Yun, Sangah Lee, Jaebum Choo* and E. K. Lee* 
기타사항 :  
Abstract

The phage display technique is a combinatorial technology in which random peptides are displayed on the surface of the phage; it is widely used to identify high-affinity peptides that bind to a target protein. However, this technique presents several problems due to non-specific binding of the phages and steric hindrance caused by blocking agents. To overcome these problems, we tested two modified methods and compared their screening performance with that of the conventional method. We used poly-His-tagged human epidermal growth factor receptor 2 (HER2) as a target protein and silica-coated magnetic particles (MPs) with an immobilized nickelnitrilotriacetic acid ligand as a solid matrix. Modified method #1 (#M1) included two negative selection steps against a blocking agent (bovine serum albumin) and nude Ni-NTA MPs, after the positive selection step using immobilized HER2 on MPs in the absence of BSA. Modified method #2 (#M2) allowed the binding of phages and HER2 in solution prior to immobilizing HER2 on the MP surface. The negative selection procedure was the same between them. The binding affinity of the phages screened by #M1 and #M2 was evaluated by phage ELISA. Two phages from #M2 (#M2-4 and 5) showed the highest binding, and between them #M2-5 was selected for affinity maturation by inserting a hairpin-structured peptide between the two #M2-5 peptides. SPR study showed the HER2-binding affinity was indeed improved by ca. 2000-fold to become comparable with that of the intact antibody (trastuzumab). The modified biopanning methods combined with the dimerization maturation can be an effective way to develop antibody-mimicking peptides for affinity binding.

Download PDF = Click Attachment, please.
List of Articles
제목 저널명 저자 기타사항

2019 Direct Visualization of a Surface-Enhanced Raman Spectroscopy Nano-Gap via E...

저널명 Applied Surface Science, in press. 

저자 Won-Hwa Park*, Sung-Gyu Park, Mijeong Kang, Moon Seop Hyun, Namhyun Choi, Dong-Ho Kim* and Jaebum Choo* 

기타사항  

2019 Application of SERS-based Microfluidics for In Vitro Diagnostics

저널명 In “Medical and Biological Applications Using Microfluidic Devices”, Book Chapter 9 (2018) in press, Springer. 

저자 Jinhyeok Jeon, Namhyun Choi, Joung-il Moon, Hao Chen and Jaebum Choo* 

기타사항 Book Chapter 

2019 Green multi-functional monomer based ion imprinted polymers for selective rem...

저널명 Journal of Colloid and Interface Science, in press. 

저자 L. Wang, J. Lia, J. Wang, X. Guo, X. Wang, J. Choo* and L. Chen* 

기타사항  

2019 SERS-based droplet microfluidics for high throughput gradient analysis file

저널명 Lab on a Chip, 19, 674-681 (2019) 

저자 J. Jeon, N. Choi, H. Chen, J. -I. Moon, L. Chen* and J. Choo* 

기타사항  

2019 High-throughput Microfluidic Imaging Flow Cytometry file

저널명 Current Opinion in Biotechnology, 55, 36-43 (2019) 

저자 Stavros Stavrakis, Gregor Holzner, Jaebum Choo* and Andrew deMello* 

기타사항  

2019 Modification of Phage Display Technique for Improved Screening of High-Aff...

저널명 Journal of Biotechnology, 289, 88-92 (2019) 

저자 Soi Yun, Sangah Lee, Jaebum Choo* and E. K. Lee* 

기타사항  


XE Login