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Humanization and directed evolution of the selenium-containing: ScFv phage abzyme
Xu Y.1; Li P.1; Nie J.1; Zhao Q.2; Guan S.1; Kuai Z.1; Qiao Y.1; Jiang X.1; Li Y.1; Li W.1; Shi Y.1; Kong W.1; Shan Y.1
2018
Source PublicationRSC Advances
ISSN20462069
Volume8Issue:31Pages:17218-17223
Abstract

According to the binding site structure and the catalytic mechanism of the native glutathione peroxidase (GPX), three glutathione derivatives, GSH-S-DNP butyl ester (hapten Be), GSH-S-DNP hexyl ester (hapten He) and GSH-S-DNP hexamethylene ester (hapten Hme) were synthesized. By a four-round panning with a human synthetic scFv phage library against three haptens, the enrichment of the scFv phage particles with specific binding activity could be determined. Three phage particles were selected binding to each glutathione derivative, respectively. After a two-step chemical mutation to convert the serine residues of the scFv phage particles into selenocysteine residues, GPX activity could be observed and determined upto 3000 U μmol in the selenium-containing scFv phage abzyme which was isolated by affinity capture against the hapten Be. Also the scFv phage abzymes elicited by different antigens displayed different catalytic activities. After a directed evolution by DNA shuffling to improve the affinity to the hapten Be, a secondary library with GPX activity was created in which the catalytic activity of the selenium-containing scFv phage abzyme could be increased 17%. This study might be helpful for new haptens or antigens design to optimize the abzymes with high binding activities and might also provide a novel scheme for GPX mimic candidates for drug development.

DOI10.1039/c8ra02798f
URLView the original
Indexed BySCI
Language英语
WOS Research AreaChemistry
WOS SubjectChemistry, Multidisciplinary
WOS IDWOS:000435822200023
Fulltext Access
Citation statistics
Document TypeJournal article
CollectionFaculty of Health Sciences
Affiliation1.Jilin University
2.Universidade de Macau
Recommended Citation
GB/T 7714
Xu Y.,Li P.,Nie J.,et al. Humanization and directed evolution of the selenium-containing: ScFv phage abzyme[J]. RSC Advances,2018,8(31):17218-17223.
APA Xu Y..,Li P..,Nie J..,Zhao Q..,Guan S..,...&Shan Y..(2018).Humanization and directed evolution of the selenium-containing: ScFv phage abzyme.RSC Advances,8(31),17218-17223.
MLA Xu Y.,et al."Humanization and directed evolution of the selenium-containing: ScFv phage abzyme".RSC Advances 8.31(2018):17218-17223.
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