phage antibody
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Author(s):  
Marjan Abri Aghdam ◽  
Mohammad Reza Tohidkia ◽  
Elham Ghamghami ◽  
Asadollah Ahmadikhah ◽  
Morteza Khanmahamadi ◽  
...  

Purpose: Production of functional recombinant antibody fragments in the periplasm of E. coli is a prerequisite step to achieve sufficient reagent for preclinical studies. Thus, the cost-effective and lab-scale production of antibody fragments demands the optimization of culture conditions. Methods: The culture conditions such as temperature, optical density (OD600) at induction, induction time, and IPTG concentration were investigated to optimize the functional expression of a phage-derived scFv molecule using a design of experiment (DoE). Additionally, the effects of different culture media and osmolyte supplements on the expression yield of scFv were examined. Results: The developed 2FI regression model indicated the significant linear effect of the incubation temperature, the induction time, and the induction OD600 on the expression yield of functional scFv. Besides, the statistical analysis indicated that two significant interactions of the temperature/induction time and the temperature/induction OD600 significantly interplay to increase the yield. Further optimization showed that the expression level of functional scFv was the most optimal when the cultivation was undertaken either in the TB medium or in the presence of media supplements of 0.5 M sorbitol or 100 mM glycine betaine. Conclusion: In the present study, for the first time, we successfully implemented DoE to comprehensively optimize the culture conditions for the expression of scFv molecules in a phage antibody display setting, where scFv molecules can be isolated from a tailor-made phage antibody library known as "Human Single Fold scFv Library I."


2020 ◽  
Vol 28 (1) ◽  
pp. 221-235
Author(s):  
Bahareh Zarei ◽  
Zahra Javidan ◽  
Elnaz Fatemi ◽  
Fatemeh Rahimi Jamnani ◽  
Shohreh Khatami ◽  
...  

2020 ◽  
Vol 56 (1) ◽  
pp. 106-113
Author(s):  
O. I. Guliy ◽  
N. S. Velichko ◽  
Yu. P. Fedonenko ◽  
V. D. Bunin

2019 ◽  
Vol 61 (11) ◽  
pp. 801-815 ◽  
Author(s):  
Rose H. Reader ◽  
Robert G. Workman ◽  
Ben C. Maddison ◽  
Kevin C. Gough

Talanta ◽  
2019 ◽  
Vol 197 ◽  
pp. 397-405 ◽  
Author(s):  
Chongxin Xu ◽  
Xiaoqin Liu ◽  
Yuan Liu ◽  
Xiao Zhang ◽  
Cunzheng Zhang ◽  
...  

PLoS ONE ◽  
2017 ◽  
Vol 12 (1) ◽  
pp. e0167860 ◽  
Author(s):  
Yurie Enomoto-Okawa ◽  
Yuka Maeda ◽  
Nozomi Harashima ◽  
Yumika Sugawara ◽  
Fumihiko Katagiri ◽  
...  

BMC Cancer ◽  
2017 ◽  
Vol 17 (1) ◽  
Author(s):  
Karen Marie Juul Sørensen ◽  
Theresa Meldgaard ◽  
Connie Jenning Melchjorsen ◽  
Agla J. Fridriksdottir ◽  
Henrik Pedersen ◽  
...  

2016 ◽  
Author(s):  
Douglas M. Templeton ◽  
Michael Schwenk ◽  
Reinhild Klein ◽  
John H. Duffus

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