Asset Details
Do you wish to reserve the book?
Expression of actively soluble antigen-binding fragment (Fab) antibody and GFP fused Fab in the cytoplasm of the engineered Escherichia coli
Hey, we have placed the reservation for you!
By the way, why not check out events that you can attend while you pick your title.
Oops! Something went wrong.
Looks like we were not able to place the reservation. Kindly try again later.
Are you sure you want to remove the book from the shelf?
Expression of actively soluble antigen-binding fragment (Fab) antibody and GFP fused Fab in the cytoplasm of the engineered Escherichia coli
Do you wish to request the book?
Expression of actively soluble antigen-binding fragment (Fab) antibody and GFP fused Fab in the cytoplasm of the engineered Escherichia coli
Please be aware that the book you have requested cannot be checked out. If you would like to checkout this book, you can reserve another copy
We have requested the book for you!
Your request is successful and it will be processed during the Library working hours. Please check the status of your request in My Requests.
Oops! Something went wrong.
Looks like we were not able to place your request. Kindly try again later.
Journal Article
Expression of actively soluble antigen-binding fragment (Fab) antibody and GFP fused Fab in the cytoplasm of the engineered Escherichia coli
2020
Overview
The expression of recombinant antibody fragments in the cytoplasmic space of
Escherichia coli
and the refolding process for restoring the structure and activity of such antibodies are not efficient. Herein, fragment antigen-binding (Fab) antibodies against miroestrol and deoxymiroestrol (MD-Fab) and their fusions with a green fluorescent protein (GFP) were expressed. The reactive MD-Fabs were successfully expressed as soluble and active forms in the cytoplasm of the SHuffle® T7
E. coli
strain. Regarding the construct of MD-Fab alone, V
H
–C
H
1 could associate V
L
–C
L
into Fab in the oxidizing cytoplasm of the
E. coli
strain, and no additional in vitro refolding was needed. In the case of the fusions with GFP, when the C-terminus of V
H
–C
H
1 was linked with the N-terminus of GFP, the MD-Fab binding reactivity was retained, but the fluorescent activity of GFP interfered. When the C-terminus of GFP was linked to the N-terminus of V
L
–C
L
, the binding activity of MD-Fab was not observed. The constructed MD-Fabs had higher specificity toward deoxymiroestrol than the parental monoclonal antibody clone 12G11. In conclusion, MD-Fabs could be expressed using SHuffle® T7
E. coli
cells. This process could be considered an economical, productive, and effective method to produce antibody fragments for immunoassay techniques.
Publisher
Springer Science and Business Media LLC,Springer Netherlands,Springer Nature B.V
Subject
/ Antigens
/ Biomedical and Life Sciences
/ E coli
/ Enzyme-Linked Immunosorbent Assay
/ Enzyme-Linked Immunosorbent Assay - methods
/ Escherichia coli - metabolism
/ Escherichia coli Proteins - genetics
/ Escherichia coli Proteins - metabolism
/ Fab
/ Immunoglobulin Fab Fragments
/ Immunoglobulin Fab Fragments - chemistry
/ Immunoglobulin Fab Fragments - genetics
/ Immunoglobulin Fab Fragments - metabolism
/ Protein Engineering - methods
/ Recombinant Fusion Proteins - genetics
