scholarly journals Fragment antigen binding domains (Fabs) as tools to study assembly-line polyketide synthases

2022 ◽  
Vol 7 (1) ◽  
pp. 506-512
Author(s):  
Katarina M. Guzman ◽  
Chaitan Khosla
Keyword(s):  
Assembly Line ◽  
Polyketide Synthases ◽  
Antigen Binding ◽  
Binding Domains ◽  
Fragment Antigen Binding

scholarly journals Characterization and validation of fragment antigen-binding (Fab) antibody-based immunoassay for deoxymiroestrol, a potent phytoestrogen from Pueraria candollei

2021 ◽  
Author(s):  
Worapol Sae-foo ◽  
Supaluk Krittanai ◽  
Wipawee Juengsanguanpornsuk ◽  
Gorawit Yusakul ◽  
Tharita Kitisripanya ◽  
...  
Keyword(s):  
Quantitative Method ◽  
Estrogenic Activity ◽  
Limit Of Detection ◽  
Hormone Replacement ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antigen Binding ◽  
Pueraria Candollei ◽  
Specific Determination ◽  
Fragment Antigen Binding

Deoxymiroestrol is the most potent phytoestrogen in chromenes group that has been found in Pueraria candollei, Thai name known as Kwao Krua Khao. Several studies reported estrogenic activity of P. candollei in order to using as hormone replacement therapy for postmenopausal women. Previously, specific determination of deoxymiroestrol content by enzyme-linked immunosorbent assay (ELISA) using polyclonal antibody (pAb) have been reported. However, production of pAb has limitation and variability from different batches. Therefore, in this study, we established quantitative method for determination of deoxymiroestrol using fragment antigen-binding (Fab) antibody-based immunoassay. The developed immunoassay has specificity to deoxymiroestrol with a calibration range of 15.6-1000 ng mL-1. Precision including intra-assay and inter-assay are 1.48-7.11 and 0.58-9.31%, respectively. Accuracy of the assay showed in recovery between 99.77-101.61% when spike deoxymiroestrol standard into the samples. The limit of detection (LOD) is 30.80 ng mL-1. Comparation antibody-based immunoassay for determination of deoxymiroestrol using Fab with pAb was represented consistency (R2 = 0.9807) when analysis roots bark of Pueraria candollei from difference areas. Therefore, this development assay can apply to determine deoxymiroestrol content in the plant samples.

Expression of functional eGFP-fused antigen-binding fragment of ranibizumab in Pichia pastoris

Bioimpacts ◽  
2021 ◽  
Author(s):  
Shirin Movaghar Asareh ◽  
Tahereh Savei ◽  
Sareh Arjmand ◽  
Seyed Omid Ranaei Siadat ◽  
Fataneh Fatemi ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Fluorescent Protein ◽  
Antibody Fragment ◽  
Binding Activity ◽  
Age Related Macular Degeneration ◽  
Light Chains ◽  
Antigen Binding ◽  
Active Structure ◽  
Age Related ◽  
Fragment Antigen Binding

Introduction: Ranibizumab is a mouse monoclonal antibody fragment antigen-binding (Fab) against human vascular endothelial growth factor-A (VEGF-A), inhibiting angiogenesis. This antibody is commercially produced in Escherichia coli host and used to treat wet age-related macular degeneration (AMD).Methods: In this study, the heavy and light chains of ranibizumab were expressed in Pichia pastoris. The expressed chains were incubated overnight at 4°C for interaction. The formation of an active structure was evaluated based on the interaction with substrate VEGF-A using an indirect ELISA, and an electrochemical setup. Furthermore, reconstruction of split enhanced green fluorescent protein (eGFP) reporter, chimerized at the C-terminus of the heavy and light chains, was used to characterize chains’ interaction. Results: P. pastoris efficiently expressed designed constructs and secreted them into the culture medium. The anti-Fab antibody detected the constructed Fab structure in western blot analysis. Reconstruction of the split reporter confirmed the interaction between heavy and light chains. The designed ELISA and electrochemical setup results verified the binding activity of the recombinant Fab structure against VEGF-A. Conclusion: In this work, we indicated that the heavy and light chains of ranibizumab Fab fragments (with or without linkage to split parts of eGFP protein) were produced in P. pastoris. The fluorescence of reconstructed eGFP was detected after incubating the equal ratio of chimeric-heavy and light chains. Immunoassay and electrochemical tests verified the bioactivity of constructed Fab. The data suggested that P. pastoris could be considered a potential efficient eukaryotic host for ranibizumab production.

scholarly journals Development of chimeric antigen receptors targeting T-cell malignancies using two structurally different anti-CD5 antigen binding domains in NK and CRISPR-edited T cell lines

2017 ◽  
Vol 7 (3) ◽  
pp. e1407898 ◽  
Author(s):  
Sunil S. Raikar ◽  
Lauren C. Fleischer ◽  
Robert Moot ◽  
Andrew Fedanov ◽  
Na Yoon Paik ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Lines ◽  
Antigen Binding ◽  
Antigen Receptors ◽  
Chimeric Antigen Receptors ◽  
Binding Domains ◽  
T Cell Lines ◽  
T Cell Malignancies

scholarly journals Enhancing Antibodies’ Binding Capacity through Oriented Functionalization of Plasmonic Surfaces

Nanomaterials ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 2620
Author(s):  
Maria Laura Coluccio ◽  
Fabiana Grillo ◽  
Valentina Onesto ◽  
Virginia Garo ◽  
Cinzia Scala ◽  
...  
Keyword(s):  
Silicon Surface ◽  
Binding Capacity ◽  
Protein A ◽  
Antigen Binding ◽  
Crucial Importance ◽  
Enhanced Fluorescence ◽  
Immobilized Antibodies ◽  
Research Fields ◽  
Fragment Antigen Binding ◽  
First Time

Protein A has long been used in different research fields due to its ability to specifically recognize immunoglobulins (Ig). The protein derived from Staphylococcus aureus binds Ig through the Fc region of the antibody, showing its strongest binding in immunoglobulin G (IgG), making it the most used protein in its purification and detection. The research presented here integrates, for the first time, protein A to a silicon surface patterned with gold nanoparticles for the oriented binding of IgG. The signal detection is conveyed through a metal enhanced fluorescence (MEF) system. Orienting immunoglobulins allows the exposition of the fragment antigen-binding (Fab) region for the binding to its antigen, substantially increasing the binding capacity per antibody immobilized. Antibodies orientation is of crucial importance in many diagnostics devices, particularly when either component is in limited quantities.

scholarly journals Foreign or Domestic CARs: Receptor Ligands as Antigen-Binding Domains

2014 ◽  
Vol 2 (1) ◽  
pp. 23-36 ◽  
Author(s):  
Donald Shaffer ◽  
Penghui Zhou ◽  
Stephen Gottschalk
Keyword(s):  
Antigen Binding ◽  
Receptor Ligands ◽  
Binding Domains

In Vitro Investigation of Crosstalk between Fatty Acid and Polyketide Synthases in the Andrimid Biosynthetic Assembly Line

ChemBioChem ◽  
2016 ◽  
Vol 17 (22) ◽  
pp. 2137-2142 ◽  
Author(s):  
Fumihiro Ishikawa ◽  
Hiroyasu Sugimoto ◽  
Hideaki Kakeya
Keyword(s):  
Fatty Acid ◽  
Assembly Line ◽  
Polyketide Synthases ◽  
In Vitro Investigation

scholarly journals CARs: Beyond T Cells and T Cell-Derived Signaling Domains

2020 ◽  
Vol 21 (10) ◽  
pp. 3525 ◽  
Author(s):  
Nico M. Sievers ◽  
Jan Dörrie ◽  
Niels Schaft
Keyword(s):  
Cell Types ◽  
Antigen Binding ◽  
Cell Type ◽  
Transfected Cells ◽  
Effector Functions ◽  
Binding Domains ◽  
Specific Effector ◽  
Different Cell Types ◽  
Intracellular Domains ◽  
Signaling Domains

When optimizing chimeric antigen receptor (CAR) therapy in terms of efficacy, safety, and broadening its application to new malignancies, there are two main clusters of topics to be addressed: the CAR design and the choice of transfected cells. The former focuses on the CAR construct itself. The utilized transmembrane and intracellular domains determine the signaling pathways induced by antigen binding and thereby the cell-specific effector functions triggered. The main part of this review summarizes our understanding of common signaling domains employed in CARs, their interactions among another, and their effects on different cell types. It will, moreover, highlight several less common extracellular and intracellular domains that might permit unique new opportunities. Different antibody-based extracellular antigen-binding domains have been pursued and optimized to strike a balance between specificity, affinity, and toxicity, but these have been reviewed elsewhere. The second cluster of topics is about the cellular vessels expressing the CAR. It is essential to understand the specific attributes of each cell type influencing anti-tumor efficacy, persistence, and safety, and how CAR cells crosstalk with each other and bystander cells. The first part of this review focuses on the progress achieved in adopting different leukocytes for CAR therapy.

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