scholarly journals FN3 Domain Displaying Double Epitopes: A Cost-Effective Strategy for Producing Substitute Antigens

2021 ◽  
Vol 8 ◽  
Author(s):  
Yao Ruan ◽  
Shuangying Chao ◽  
Xuejun Hu ◽  
Longzhen Lu ◽  
Yue Lin ◽  
...  
Keyword(s):  
Expression System ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Effective Strategy ◽  
Scanning Calorimetry ◽  
Specific Antibodies ◽  
Peptide Epitopes ◽  
Cost Effective Strategy ◽  
Loop Region ◽  
C Terminus

Construction of substitute antigens based on alternative scaffold proteins is a promising strategy in bioassay technology. In this study, we proposed a strategy for constructing substitute antigens derived from 10th human fibronectin type III (FN3) using two peptide epitopes of terminal pro-brain natriuretic peptide (NT-proBNP) as an example. The base sequences encoding the two antigenic epitopes of NT-proBNP were recombined into the FG loop region and the C-terminus of FN3, fused by 4 GS or polyN linker. The fusion proteins (named FN3-epitopes-4GS and FN3-epitopes-polyN, respectively) were expressed and purified cost-effectively using an Escherichia coli expression system. The immunoreactivity of recombinant substitutes was preliminarily confirmed by western blot analysis using epitope-specific antibodies. The sandwich enzyme-linked immunosorbent assay demonstrated that either FN3-epitopes-polyN or FN3-epitopes-4GS was highly sensitive, and FN3-epitopes-polyN exhibited better kinetics to specific antibodies than FN3-epitopes-4GS, showing a linear dose-response relationship in the concentration range of 0.06–12.85 ng/ml, which suggest that the polyN linker was more suitable for constructing the FN3-based substitute antigens compared to the 4 GS linker. Furthermore, the serum stability test and differential scanning calorimetry analysis showed that the recombinant FN3-epitopes-polyN maintained the original stability of FN3. Therefore, it was confirmed that FN3 could be engineered to construct a stable biomacromolecular substitute for displaying double epitopes of antigen proteins, such as NT-proBNP. In summary, a cost-effective strategy to produce NT-proBNP substitute antigens with good immunoreactivity and physicochemical stability was established in this work, which may provide potential uses for the production of other substitute antigens in the future.

A cost effective strategy to fabricate STA@PF@Cu2O hierarchical structure on wood surface: aimed at superhydrophobic modification

2021 ◽  
Vol 55 (2) ◽  
pp. 565-583
Author(s):  
Ke Zhan ◽  
Quanxiong Lu ◽  
Sengwei Xia ◽  
Congnan Guo ◽  
Sisi Zhao ◽  
...  
Keyword(s):  
Hierarchical Structure ◽  
Wood Surface ◽  
Cost Effective ◽  
Effective Strategy ◽  
Cost Effective Strategy

A cost effective strategy for managing GI bleeding secondary to peptic ulcer disease

1995 ◽  
Vol 108 (4) ◽  
pp. A18 ◽  
Author(s):  
I Hussain ◽  
WH Farrar ◽  
EJ Sofian ◽  
TF Bader ◽  
JD Strom ◽  
...  
Keyword(s):  
Peptic Ulcer ◽  
Peptic Ulcer Disease ◽  
Cost Effective ◽  
Effective Strategy ◽  
Gi Bleeding ◽  
Ulcer Disease ◽  
Cost Effective Strategy

scholarly journals Development of an Immunoassay for Detection of Staphylococcal Enterotoxin-Like J, A Non-Characterized Toxin

Toxins ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 458 ◽  
Author(s):  
Hisaya Ono ◽  
Nobuaki Hachiya ◽  
Yasunori Suzuki ◽  
Ikunori Naito ◽  
Shouhei Hirose ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Expression System ◽  
Sandwich Elisa ◽  
Food Poisoning ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
E Coli ◽  
Detection Systems ◽  
C Terminus ◽  
Culture Supernatants

Staphylococcal enterotoxins (SEs) are the cause of staphylococcal food poisoning (SFP) outbreaks. Recently, many new types of SEs and SE-like toxins have been reported, but it has not been proved whether these new toxins cause food poisoning. To develop an immunoassay for detection of SE-like J (SElJ), a non-characterized toxin in SFP, a mutant SElJ with C-terminus deletion (SElJ∆C) was expressed and purified in an E. coli expression system. Anti-SElJ antibody was produced in rabbits immunized with the SElJ∆C. Western blotting and sandwich enzyme-linked immunosorbent assay (ELISA) detection systems were established and showed that the antibody specifically recognizes SElJ without cross reaction to other SEs tested. The limit of detection for the sandwich ELISA was 0.078 ng/mL, showing high sensitivity. SElJ production in S. aureus was detected by using the sandwich ELISA and showed that selj-horboring isolates produced a large amount of SElJ in the culture supernatants, especially in that of the strain isolated from a food poisoning outbreak in Japan. These results demonstrate that the immunoassay for detection of SElJ is specific and sensitive and is useful for determining the native SElJ production in S. aureus isolated from food poisoning cases.

scholarly journals Scaling-Up the Recognition of Indigenous and Community Land Rights: Opportunities, Costs and Climate Implications

2021 ◽  
Author(s):  
◽  
Keyword(s):  
Indigenous Peoples ◽  
Cost Effective ◽  
Land Rights ◽  
Scaling Up ◽  
Local Communities ◽  
Effective Strategy ◽  
Cost Effective Strategy ◽  
International Funding ◽  
Growing Body ◽  
Alternative Solutions

A growing body of evidence suggests that recognition of the collective tenure rights of Indigenous Peoples, local communities, and Afro-descendants is a powerful and cost-effective strategy for addressing the climate and biodiversity crises. In spite of this, international funding for rights recognition pales in comparison to donor mobilization around alternative solutions to these crises.

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