Production of Stabilized scFv Antibody Fragments in the E. coli Bacterial Cytoplasm

2014 ◽  
pp. 171-184 ◽  
Author(s):  
Lilach Vaks ◽  
Itai Benhar
Keyword(s):  
Antibody Fragments ◽  
Scfv Antibody ◽  
E Coli ◽  
Bacterial Cytoplasm

scholarly journals Chaperone-Assisted Soluble Expression of a Humanized Anti-EGFR ScFv Antibody in E. Coli

2015 ◽  
Vol 5 (Suppl 1) ◽  
pp. 621-627 ◽  
Author(s):  
Kamal Veisi ◽  
Safar Farajnia ◽  
Nosratollah Zarghami ◽  
Hamid Reza Khoram Khorshid ◽  
Nasser Samadi ◽  
...  
Keyword(s):  
Soluble Expression ◽  
Scfv Antibody ◽  
E Coli

scholarly journals Making Soup: Preparing and Validating Molecular Simulations of the Bacterial Cytoplasm

2019 ◽  
Author(s):  
Leandro Oliveira Bortot ◽  
Zahedeh Bashardanesh ◽  
David van der Spoel
Keyword(s):  
Large Scale ◽  
Biological Properties ◽  
Computational Power ◽  
E Coli ◽  
Solvent Models ◽  
Explicit Solvent ◽  
Computational Modeling And Simulation ◽  
Dynamics Simulations ◽  
The Individual ◽  
Bacterial Cytoplasm

Biomolecular crowding affects the biophysical and biochemical behavior of macro- molecules when compared to the dilute environment present in experiments made with isolated proteins. Computational modeling and simulation are useful tools to study how crowding affects the structural dynamics and biological properties of macromolecules. As computational power increased, modeling and simulating large scale all-atom explicit solvent models of the prokaryote cytoplasm become possible. In this work, we build an atomistic model of the cytoplasm of Escherichia coli composed of 1.5 million atoms and submit it to a total of 3 μs of molecular dynamics simulations. The properties of biomolecules under crowding conditions are compared to those from simulations of the individual compounds under dilute conditions. The simulation model is found to be consistent with experimental data about the diffusion coefficient and stability of macromolecules under crowded conditions. In order to stimulate further work we provide a Python script and a set of files that enables other researchers to build their own E. coli cytoplasm models to address questions related to crowding.<br>

scholarly journals Systematic screening of soluble expression of antibody fragments in the cytoplasm of E. coli

2016 ◽  
Vol 15 (1) ◽  
Author(s):  
Anna Gaciarz ◽  
Johanna Veijola ◽  
Yuko Uchida ◽  
Mirva J. Saaranen ◽  
Chunguang Wang ◽  
...  
Keyword(s):  
Antibody Fragments ◽  
Soluble Expression ◽  
Systematic Screening ◽  
E Coli

A new expression vector facilitating production and functional analysis of scFv antibody fragments selected from Tomlinson I + J phagemid libraries

2015 ◽  
Vol 167 (2) ◽  
pp. 95-102 ◽  
Author(s):  
Karolina Ossysek ◽  
Tomasz Uchański ◽  
Małgorzata Kulesza ◽  
Monika Bzowska ◽  
Tomasz Klaus ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Expression Vector ◽  
Antibody Fragments ◽  
Scfv Antibody

scholarly journals Selection of scFv Antibody Fragments Binding to Human Blood versus Lymphatic Endothelial Surface Antigens by Direct Cell Phage Display

PLoS ONE ◽  
2015 ◽  
Vol 10 (5) ◽  
pp. e0127169 ◽  
Author(s):  
Thomas Keller ◽  
Romana Kalt ◽  
Ingrid Raab ◽  
Helga Schachner ◽  
Corina Mayrhofer ◽  
...  
Keyword(s):  
Phage Display ◽  
Human Blood ◽  
Surface Antigens ◽  
Antibody Fragments ◽  
Scfv Antibody ◽  
Endothelial Surface ◽  
Direct Cell ◽  
Selection Of

scholarly journals Dynamic Characteristics of Allosteric Pathways in scFv Antibody Fragments

2015 ◽  
Vol 108 (2) ◽  
pp. 377a
Author(s):  
Amit Srivastava ◽  
Malgorzata B. Tracka ◽  
Shahid Uddin ◽  
Jose Casas-Finet ◽  
Dennis R. Livesay ◽  
...  
Keyword(s):  
Dynamic Characteristics ◽  
Antibody Fragments ◽  
Scfv Antibody

scholarly journals Specific Secretion of Active Single-Chain Fv Antibodies into the Supernatants of Escherichia coliCultures by Use of the Hemolysin System

2000 ◽  
Vol 66 (11) ◽  
pp. 5024-5029 ◽  
Author(s):  
Luis A. Fernández ◽  
Isabel Sola ◽  
Luis Enjuanes ◽  
Víctor de Lorenzo
Keyword(s):  
Escherichia Coli ◽  
Transport System ◽  
Culture Media ◽  
Single Chain ◽  
Extracellular Medium ◽  
Simple Method ◽  
E Coli ◽  
Single Chain Fv ◽  
Bacterial Cytoplasm ◽  
Culture Supernatants

ABSTRACT A simple method for the nontoxic, specific, and efficient secretion of active single-chain Fv antibodies (scFvs) into the supernatants ofEscherichia coli cultures is reported. The method is based on the well-characterized hemolysin transport system (Hly) of E. coli that specifically secretes the target protein from the bacterial cytoplasm into the extracellular medium without a periplasmic intermediate. The culture media that accumulate these Hly-secreted scFv's can be used in a variety of immunoassays without purification. In addition, these culture supernatants are stable over long periods of time and can be handled basically as immune sera.

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