scholarly journals Strategies and Applications of Antigen-Binding Fragment (Fab) Production in Escherichia coli

2021 ◽  
Vol 03 (02) ◽  
pp. e39-e49
Author(s):  
Hui Chen ◽  
Jun-Sheng Chen ◽  
Pameila Paerhati ◽  
Tanja Jakos ◽  
Si-Yi Bai ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antigen Binding ◽  
Expression Systems ◽  
Single Chain ◽  
Therapeutic Modalities ◽  
Fab Fragments ◽  
Single Chain Variable Fragments ◽  
Escherichia Coli Expression ◽  
Fermentation Technology ◽  
Shed Light

AbstractWith the advancement of genetic engineering, monoclonal antibodies (mAbs) have made far-reaching progress in the treatment of various human diseases. However, due to the high cost of production, the increasing demands for antibody-based therapies have not been fully met. Currently, mAb-derived alternatives, such as antigen-binding fragments (Fab), single-chain variable fragments, bispecifics, nanobodies, and conjugated mAbs have emerged as promising new therapeutic modalities. They can be readily prepared in bacterial systems with well-established fermentation technology and ease of manipulation, leading to the reduction of overall cost. This review aims to shed light on the strategies to improve the expression, purification, and yield of Fab fragments in Escherichia coli expression systems, as well as current advances in the applications of Fab fragments.

scholarly journals Single Chain Variable Fragments Produced in Escherichia coli against Heat-Labile and Heat-Stable Toxins from Enterotoxigenic E. coli

PLoS ONE ◽  
2015 ◽  
Vol 10 (7) ◽  
pp. e0131484 ◽  
Author(s):  
Christiane Y. Ozaki ◽  
Caio R. F. Silveira ◽  
Fernanda B. Andrade ◽  
Roberto Nepomuceno ◽  
Anderson Silva ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Single Chain ◽  
E Coli ◽  
Heat Stable ◽  
Heat Labile ◽  
Single Chain Variable Fragments

scholarly journals Comparison of Gene Expression Between Two Types of Anti-EGFRvIII ScFv Antibodies Having Different Variable Domain Orders in Escherichia coli

2017 ◽  
Vol 21 (1) ◽  
pp. 29
Author(s):  
Kartika Sari Dewi ◽  
Asrul Muhamad Fuad
Keyword(s):  
Escherichia Coli ◽  
Expression System ◽  
Binding Activity ◽  
Extracellular Proteins ◽  
Antigen Binding ◽  
Dna Encoding ◽  
Single Chain ◽  
Colony Pcr ◽  
E Coli ◽  
Variable Domains

Several studies reported that the expression of various kinds of Single-chain variable fragment (scFv) antibodies in Escherichia coli are significantly influenced by the order of their variable domains. To date, the effect of the order of variable domains in the expression of scFv antibodies against epidermal growth factor receptor variant III (EGFRvIII) has not been reported. This study aimed to compare the expression between VH-linker-VL and VL-linker-VH domain orders of the anti-EGFRvIII scFv antibodies in E. coli expression system. Recombinant plasmids inserted with DNA encoding scFv proteins were transformed into E. coli NiCo21(DE3) competent cells and characterized by colony PCR. The expression of scFv proteins was done by using optimum concentration of inducer. Total proteins, soluble periplasmic and cytoplasmic proteins, also extracellular proteins were isolated, subsequently characterized by SDS-PAGE, Slot Blot, and ImageJ software analyses. The antigen-binding activity of both scFvs proteins against EGFRvIII was observed. The results showed that the relative percentage of scFv expression with VH-linker-VL domain order is higher than that of VL-linker-VH in each compartment. Moreover, both of scFvs proteins have antigen-binding activity against EGFRvIII.

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