TROUBLESHOOTING IN EXPRESSION AND PURIFICATION OF RECOMBINANT SEVERE ACUTE RESPIRATORY SYNDROME-ASSOCIATED CORONAVIRUS NUCLEOCAPSID PROTEIN IN Escherichia coli BL21

The thermostability and catalytic activity of endoglucanase from thermophilic Thermotoga maritima were improved by evolutionary molecular engineering. The cel12B gene from Thermotoga maritima was cloned and expressed, and the expressed Cel12B protein was purified by His-tag affinity chromatography. The constructed plasmids were screened on LB plate with ampicillin using Escherichia coli BL21 (DE3) as a host. The results showed that both the enzyme activity and yield of the Cell12B protein expressed in pET15b-cel12B were 2 times as that of the protein expressed in pET20b-cel12B.

Download Full-text

Naturally Occurring Anti-Escherichia coli Protein Antibodies in the Sera of Healthy Humans Cause Analytical Interference in a Recombinant Nucleocapsid Protein-Based Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Severe Acute Respiratory Syndrome

Clinical and Vaccine Immunology ◽

10.1128/cvi.00136-06 ◽

2006 ◽

Vol 14 (1) ◽

pp. 99-101 ◽

Cited By ~ 8

Author(s):

Chi Wai Yip ◽

Chung Chau Hon ◽

Fanya Zeng ◽

Ken Y. C. Chow ◽

Kwok Hung Chan ◽

...

Keyword(s):

Escherichia Coli ◽

Severe Acute Respiratory Syndrome ◽

Immunosorbent Assay ◽

Nucleocapsid Protein ◽

False Positives ◽

Enzyme Linked Immunosorbent Assay ◽

Healthy Humans ◽

Analytical Interference ◽

Human Sera ◽

Recombinant Nucleocapsid Protein

ABSTRACT We reported the analytical interference of anti-Escherichia coli protein (EP) antibodies in human sera and residual EP in a recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay as a possible source of false positives in severe acute respiratory syndrome serodiagnosis. The rate of false positives was significantly reduced by adding mouse anti-EP antiserum in the blocking step.

Download Full-text

Enhancing the production of γ-aminobutyric acid in Escherichia coli BL21 by engineering the enzymes of the regeneration pathway of the coenzyme factor pyridoxal 5’-phosphate

World Journal of Microbiology and Biotechnology ◽

10.1007/s11274-021-03103-5 ◽

2021 ◽

Vol 37 (8) ◽

Author(s):

Ping Yu ◽

Jian Ma ◽

Pengzhi Zhu ◽

Qingwei Chen ◽

Qili Zhang

Keyword(s):

Escherichia Coli ◽

Aminobutyric Acid ◽

Escherichia Coli Bl21 ◽

Regeneration Pathway

Download Full-text

Immunological imprinting of the antibody response in COVID-19 patients

Nature Communications ◽

10.1038/s41467-021-23977-1 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Teresa Aydillo ◽

Alexander Rombauts ◽

Daniel Stadlbauer ◽

Sadaf Aslam ◽

Gabriela Abelenda-Alonso ◽

...

Keyword(s):

Immune Response ◽

Severe Acute Respiratory Syndrome ◽

Antibody Response ◽

Humoral Immune Response ◽

Nucleocapsid Protein ◽

Spike Protein ◽

Ongoing Research ◽

Variable Regions ◽

Humoral Immune ◽

Human Coronaviruses

AbstractIn addition to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), humans are also susceptible to six other coronaviruses, for which consecutive exposures to antigenically related and divergent seasonal coronaviruses are frequent. Despite the prevalence of COVID-19 pandemic and ongoing research, the nature of the antibody response against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is unclear. Here we longitudinally profile the early humoral immune response against SARS-CoV-2 in hospitalized coronavirus disease 2019 (COVID-19) patients and quantify levels of pre-existing immunity to OC43, HKU1 and 229E seasonal coronaviruses, and find a strong back-boosting effect to conserved but not variable regions of OC43 and HKU1 betacoronaviruses spike protein. However, such antibody memory boost to human coronaviruses negatively correlates with the induction of IgG and IgM against SARS-CoV-2 spike and nucleocapsid protein. Our findings thus provide evidence of immunological imprinting by previous seasonal coronavirus infections that can potentially modulate the antibody profile to SARS-CoV-2 infection.

Download Full-text