Selection of single-chain variable fragment antibodies against fenitrothion by ribosome display

2012 ◽  
Vol 421 (1) ◽  
pp. 130-137 ◽  
Author(s):  
Yihui Luo ◽  
Yuxian Xia
Keyword(s):  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Ribosome Display ◽  
Selection Of

Selection of a Single Chain Variable Fragment Antibody against Ivermectin from a Phage Displayed Library

2010 ◽  
Vol 58 (9) ◽  
pp. 5387-5391 ◽  
Author(s):  
Shuang Wen ◽  
Xiao Zhang ◽  
Yuan Liu ◽  
Qingqing Zhang ◽  
Xianjin Liu ◽  
...  
Keyword(s):  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Selection Of

Selection of Single Chain Variable Fragments Specific for the Human-Inducible Costimulator Using Ribosome Display

2012 ◽  
Vol 168 (5) ◽  
pp. 967-979 ◽  
Author(s):  
Yangbin Pan ◽  
Weiping Mao ◽  
Xuanxuan Liu ◽  
Chong Xu ◽  
Zhijuan He ◽  
...  
Keyword(s):  
Single Chain ◽  
Ribosome Display ◽  
Inducible Costimulator ◽  
Single Chain Variable Fragments ◽  
Selection Of

scholarly journals Generation and Selection of a Panel of Pan-Filovirus Single-Chain Antibodies using Cell-Free Ribosome Display

2018 ◽  
Author(s):  
Adinarayana Kunamneni ◽  
Elizabeth C. Clarke ◽  
Chunyan Ye ◽  
Steven B. Bradfute ◽  
Ravi Durvasula
Keyword(s):  
Hemorrhagic Fever ◽  
Cross Reactivity ◽  
Free Ribosome ◽  
Rapid Diagnostics ◽  
Single Chain ◽  
Ribosome Display ◽  
Single Chain Antibodies ◽  
Single Chain Fv ◽  
New Generation ◽  
Selection Of

AbstractFiloviruses, which include ebolaviruses and marburgvirus, can cause outbreaks of highly lethal hemorrhagic fever. This disease causes significant morbidity and mortality in humans and non-human primates, with human fatality rates reaching 90% during some outbreaks. Currently, there are a lack of licensed vaccines or antivirals for these viruses. Since early symptoms of filovirus infection mimic more common diseases, there is a strong unmet public health and biodefense need for broad-spectrum filovirus rapid diagnostics. We have generated a panel of mouse single-chain Fv-antibodies (scFvs) to filovirus glycoproteins (GPs) using cell-free ribosome display and determined their cross-reactivity profiles to all known filovirus species. Two scFvs (4-2 and 22-1) were able to detect all known Ebolavirus and Marburgvirus species. This is the first report on ribosome display scFvs that can detect a broad set of filovirus GPs, which demonstrates their potential use in the development of a new generation of rapid diagnostic immunoassays.

scholarly journals A Cell-Based Subtractive Panning Strategy for Selection of Conformation-Specific Single-Chain Variable-Fragment (scFv) against Dimerization Domain of EGFR

2021 ◽  
Author(s):  
Reza Valadan ◽  
Mina Dabiri ◽  
Mohsen Tehrani ◽  
Gholamreza Hashemi Tabar ◽  
Alireza Rafiei
Keyword(s):  
Inhibition Test ◽  
Functional Domain ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Dimerization Domain ◽  
Antitumor Effects ◽  
Egfr Inhibition ◽  
A Cell ◽  
Cell Elisa ◽  
Selection Of

Abstract Background and Objective: Overexpression of the EGFR, from the ErbB receptor family, has been observed in several cancers and causes resistance to therapeutic antibodies, such as Herceptin. In this study, we produced a recombinant single-chain variable fragment (scFv) antibody against the EGFR dimerization domain. Methods: The recombinant scFv was generated using a cell-based subtractive panning strategy. Subtractive panning was performed on genetically engineered VERO/EGFR cells and cancerous MDA-MB-468 cells. Phage cell-ELISA was used to monitor the binding of the selected scFvs to the dimerization domain of EGFR. Inhibition of EGFR and HER2 dimerization by the produced scFvs were finally evaluated using the dimerization inhibition test. Results: PCR fingerprinting results showed a uniform digestion pattern following the third round of panning that confirmed the success of subtractive panning. Moreover, cell-ELISA validated the reactivity of the produced scFvs to EGFR after stimulation with EGF. The dimerization inhibition test showed the capacity of the scFvs to inhibit EGFR and HER2 dimerization. Conclusions: Directed HER2 targeting showed to be more effective to target the functional domain of the cell receptor for the complete blockade of the intracellular signaling pathway. The subtractive panning strategy used in this study could control the process of directed selection of specific antibodies against the dimerization domain of EGFR. The selected antibodies might then be functionally tested for antitumor effects in both in vitro and in vivo studies.

scholarly journals Ribosome display and selection of single-chain variable fragments effectively inhibit growth and progression of microspheres in vitro and in vivo

2018 ◽  
Vol 109 (5) ◽  
pp. 1503-1512
Author(s):  
Shangke Huang ◽  
Lu Feng ◽  
Gaili An ◽  
Xiaojin Zhang ◽  
Zixuan Zhao ◽  
...  
Keyword(s):  
Single Chain ◽  
Ribosome Display ◽  
Single Chain Variable Fragments ◽  
Selection Of

Selection of a Single Chain Variable Fragment Antibody (scFv) against Subtilisin BRC and its Interaction with Subtilisin BRC

2019 ◽  
Vol 8 (1) ◽  
pp. 24-31
Author(s):  
Chol-Jin Kim ◽  
Sunll Choe ◽  
Kum-Chol Ri ◽  
Chol-Ho Kim ◽  
Hyon-Gwang Li ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Phage Display ◽  
Affinity Purification ◽  
Phage Library ◽  
Amino Acid Residues ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
E Coli ◽  
Selection Of

Background: The focus of this study was the selection of a single chain variable fragment antibody (scFv) against subtilisin BRC, a fibrinolytic enzyme using phage display, and to characterize the interaction between the antibody and subtilisin BRC. Methods: The subtilisin BRC-specific phage clones were selected using Griffin.1 scFv phage library and sequenced. The gene of subtilisin BRC-specific scFv (scFv-BRC) from selected phage clone was expressed in E. coli and scFv-BRC was characterized. Molecular modeling of the three-dimensional (3D) structures of scFv-BRC was performed using MODELLER 9.19 modeling software and assessed by PROCHEK. Molecular docking of subtilisin BRC with scFv-BRC was carried out using PATCHDOCK. Results: The size of scFv-BRC gene is 635bp and it consists of 54bp of heavy chain region (VH), 336bp of light chain region (VL), 45bp of a linker. scFv-BRC was actively expressed by E. coli expression vector pET28a-scFv in E. coli BL21 (DE3), and the amount of expressed scFv-BRC was about 50 mg/L. Its molecular weight is ~26kDa. The CDR domain of scFv-BRC consists of 6 amino acids in CDR L1, 3 amino acids in CDR L2 and 9 amino acids in CDR L3. Docking results of subtilisin BRC and scFv-BRC showed global energy of - 56.29 kJ/mol. Furthermore, the results showed that amino acid residues in subtilisin BRC for binding with scFv-BRC are Tyr6, Ser182, Ser204, and Gln206. Conclusion: scFv against subtilisin BRC selected using phage display showed relatively strong binding energy with subtilisin BRC. The amino acid residues in subtilisin BRC for binding with scFv-BRC are not relevant to that in subtilisin BRC for binding with its substrates. These results suggested that scFv-BRC can be used as a ligand for detection and affinity purification of subtilisin BRC.

scholarly journals Selection of Diethylstilbestrol-Specific Single-Chain Antibodies from a Non-Immunized Mouse Ribosome Display Library

PLoS ONE ◽  
2012 ◽  
Vol 7 (3) ◽  
pp. e33186 ◽  
Author(s):  
Yanan Sun ◽  
Baoan Ning ◽  
Ming Liu ◽  
Xianjun Gao ◽  
Xianjun Fan ◽  
...  
Keyword(s):  
Single Chain ◽  
Ribosome Display ◽  
Single Chain Antibodies ◽  
Selection Of

Selection of single chain fragments against the phytopathogen Xanthomonas axonopodis pv. citri by ribosome display

2007 ◽  
Vol 41 (3) ◽  
pp. 383-389 ◽  
Author(s):  
Qing Yuan ◽  
Yuxian Xia ◽  
Siji Nian ◽  
Youping Yin ◽  
Yueqing Cao ◽  
...  
Keyword(s):  
Single Chain ◽  
Ribosome Display ◽  
Xanthomonas Axonopodis ◽  
Selection Of

scholarly journals A Cell-Based Subtractive Panning Strategy for Selection of Conformation-Specific Single-Chain Variable-Fragment (Scfv) Against Dimerization Domain of EGFR

2021 ◽  
Author(s):  
Reza Valadan ◽  
Mina Dabiri ◽  
Mohsen Tehrani ◽  
Gholamreza Hashemi Tabar ◽  
Alireza Rafiei
Keyword(s):  
Inhibition Test ◽  
Functional Domain ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Dimerization Domain ◽  
Antitumor Effects ◽  
Egfr Inhibition ◽  
A Cell ◽  
Cell Elisa ◽  
Selection Of

Abstract Background and Objective: Overexpression of the EGFR, from the ErbB receptor family, has been observed in several cancers and causes resistance to therapeutic antibodies, such as Herceptin. In this study, we produced a recombinant single-chain variable fragment (scFv) antibody against the EGFR dimerization domain. Methods The recombinant scFv was generated using a cell-based subtractive panning strategy. Subtractive panning was performed on genetically engineered VERO/EGFR cells and cancerous MDA-MB-468 cells. Phage cell-ELISA was used to monitor the binding of the selected scFvs to the dimerization domain of EGFR. Inhibition of EGFR and HER2 dimerization by the produced scFvs were finally evaluated using the dimerization inhibition test. Results PCR fingerprinting results showed a uniform digestion pattern following the third round of panning that confirmed the success of subtractive panning. Moreover, cell-ELISA validated the reactivity of the produced scFvs to EGFR after stimulation with EGF. The dimerization inhibition test showed the capacity of the scFvs to inhibit EGFR and HER2 dimerization. Conclusions Directed HER2 targeting showed to be more effective to target the functional domain of the cell receptor for the complete blockade of the intracellular signaling pathway. The subtractive panning strategy used in this study could control the process of directed selection of specific antibodies against the dimerization domain of EGFR. The selected antibodies might then be functionally tested for antitumor effects in both in vitro and in vivo studies.

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