Optimization of Tris/EDTA/Sucrose (TES) periplasmic extraction for the recovery of functional scFv antibodies

AMB Express
Elham GhamghamiBehzad Baradaran

Abstract

Single-chain variable fragments (scFvs) have gained increased attention among researchers in both academic and industrial fields owing to simple production in E. coli. The E. coli periplasm has been the site of choice for the expression of scFv molecules due to its oxidizing milieu facilitating correctly formation of disulfide bonds. Hence, the recovery of high-yield and biologically active species from the periplasmic space is a critical step at beginning of downstream processing. TES (Tris/EDTA/Sucrose) as a simple and efficient extraction method has been frequently used but under varied extraction conditions, over literature. This study, for the first time, aimed to interrogate the effects of four independent variables (i.e., Tris-HCl concentration, buffer's pH, EDTA concentration, and incubation time) and their potential interactions on the functional extraction yield of an scFv antibody from the periplasmic space of E. coli. The results indicated that the Tris-HCl concentration and pH are the most significant variables in the TES method and displayed a positive effect at their lower values on the functional extraction yield. Besides, the statistical analysis revealed 4 significant interactions between different variables. ...Continue Reading

References

Jan 1, 1979·The International Journal of Biochemistry·I R Beacham
Oct 14, 1964·Biochemical and Biophysical Research Communications·H C Neu, L A Heppel
Oct 4, 2003·Biotechnology Progress·Anurag S RathoreDavid E Steinmeyer
Oct 11, 2003·Journal of Biochemistry and Molecular Biology·Yu-Ching SuSheila Nathan
Oct 12, 2010·Journal of Proteome Research·Marcus TheinDirk Linke
Feb 4, 2012·Nature Protocols·Romain RouetDaniel Christ
Apr 5, 2012·Clinical & Developmental Immunology·Zuhaida Asra AhmadMuhajir Hamid
Jan 10, 2013·Methods in Molecular Biology·Shu QuanJames C A Bardwell
Sep 19, 2015·Critical Reviews in Biotechnology·Sanjeev K Gupta, Pratyoosh Shukla
Apr 27, 2016·Journal of Theoretical Biology·Mozafar MohammadiNeda Zarei
Jul 9, 2016·Critical Reviews in Microbiology·Sanjeev Kumar Gupta, Pratyoosh Shukla
Oct 28, 2017·Frontiers in Immunology·Katerina T XenakiPaul M P van Bergen En Henegouwen
Nov 22, 2017·BioImpacts : BI·Ali Mesgari-Shadi, Mohammad Hossein Sarrafzadeh
Jun 5, 2018·Applied Microbiology and Biotechnology·Farnaz FahimiYadollah Omidi
Jun 28, 2018·Frontiers in Pharmacology·Arun K DangiPratyoosh Shukla
Feb 28, 2019·Applied Microbiology and Biotechnology·Mehdi FouladiYadollah Omidi
Oct 24, 2019·Applied Microbiology and Biotechnology·Marloes L C PetrusMenzo Havenga

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BETA
transgenic
antibody display
ELISA

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ProtParam

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