Background: DynE8 is an iterative polyketide synthase (PKS) that assembles polyketide intermediates from acetate units derived from malonyl-CoA. Results: We report the first acyltransferase (AT DYN10 ) crystal structure for an iterative PKS. Conclusion: AT DYN10 protects the malonyl-enzyme, but not the acetyl-enzyme intermediate, from hydrolysis and facilitates the transfer of malonyl to the acyl carrier protein.Significance: This differs from the dual specificity exhibited by acyltransferases of mammalian FAS and other iterative PKSs.
Background
Neutralizing antibodies (NAbs) prevent pathogens from infecting host cells. Detection of SARS-CoV-2 NAbs is critical to evaluate herd immunity and monitor vaccine efficacy against SARS-CoV-2, the virus that causes COVID-19. All currently available NAb tests are lab-based and time-intensive.
Method
We develop a 10 min cellulose pull-down test to detect NAbs against SARS-CoV-2 from human plasma. The test evaluates the ability of antibodies to disrupt ACE2 receptor—RBD complex formation. The simple, portable, and rapid testing process relies on two key technologies: (i) the vertical-flow paper-based assay format and (ii) the rapid interaction of cellulose binding domain to cellulose paper.
Results
Here we show the construction of a cellulose-based vertical-flow test. The developed test gives above 80% sensitivity and specificity and up to 93% accuracy as compared to two current lab-based methods using COVID-19 convalescent plasma.
Conclusions
A rapid 10 min cellulose based test has been developed for detection of NAb against SARS-CoV-2. The test demonstrates comparable performance to the lab-based tests and can be used at Point-of-Care. Importantly, the approach used for this test can be easily extended to test RBD variants or to evaluate NAbs against other pathogens.
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