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- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2017.1
- 수록면
- 1 - 9 (9page)
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Phage display technology provides a powerful tool to screen a library for a binding molecule via an enrichment process. It has been adopted as a critical technology in the development of therapeutic antibodies. However, a major drawback of phage display technology is that because the degree of the enrichment cannot be controlled during the bio-panning process, it frequently results in a limited number of clones. In this study, we applied next-generation sequencing (NGS) to screen clones from a library and determine whether a greater number of clones can be identified using NGS than using conventional methods. Three chicken immune single-chain variable fragment (scFv) libraries were subjected to bio-panning on prostate-specific antigen (PSA). Phagemid DNA prepared from the original libraries as well as from the Escherichia coli pool after each round of bio-panning was analyzed using NGS, and the heavy chain complementarity-determining region 3 (HCDR3) sequences of the scFv clones were determined. Subsequently, through two-step linker PCR and cloning, the entire scFv gene was retrieved and analyzed for its reactivity to PSA in a phage enzyme immunoassay. After four rounds of bio-panning, the conventional colony screening method was performed for comparison. The scFv clones retrieved from NGS analysis included all clones identified by the conventional colony screening method as well as many additional clones. The enrichment of the HCDR3 sequence throughout the bio-panning process was a positive predictive factor for the selection of PSA-reactive scFv clones.
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참고문헌 (24)
참고문헌 신청
Barbas CF / 2001 / Phage Display : a Laboratory Manual / CSHL Press
Bazan J / 2012 / Phage display?a powerful technique for immunotherapy: 1. Introduction and potential of therapeutic applications / Hum Vaccin Immunother 8 : 1817 ~ 1828
Brock G / 2008 / clValid, an R package for cluster validation / J Stat Softw 25 : 1 ~ 22
Carmen S / 2002 / Concepts in antibody phage display / Brief Funct Genomic Proteomic 1 : 189 ~ 203
Chan CE / 2014 / The role of phage display in therapeutic antibody discovery / Int Immunol 26 : 649 ~ 657
Bazan J / 2012 / Phage display?a powerful technique for immunotherapy: 1. Introduction and potential of therapeutic applications / Hum Vaccin Immunother 8 : 1817 ~ 1828
Brock G / 2008 / clValid, an R package for cluster validation / J Stat Softw 25 : 1 ~ 22
Carmen S / 2002 / Concepts in antibody phage display / Brief Funct Genomic Proteomic 1 : 189 ~ 203
Chan CE / 2014 / The role of phage display in therapeutic antibody discovery / Int Immunol 26 : 649 ~ 657
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