recombinant hace2 protein  (New England Biolabs)

Journal: Vaccines

Article Title: Surrogate Virus Neutralisation Test Based on Nanoluciferase-Tagged Antigens to Quantify Inhibitory Antibodies against SARS-CoV-2 and Characterise Omicron-Specific Reactivity in a Vaccination Cohort

doi: 10.3390/vaccines11121832

Figure Lengend Snippet: Detection of inhibitory antibodies by sVNT. ( A ) In-house sVNT employing the secNLuc-RBD construct was used to determine antibodies inhibiting binding of the RBD to hACE2 in vaccinee sera at time points t0–t5. The cut-off determined by ROC curve analysis (25% inhibition) is indicated by the dashed line. Horizontal bars delineate the variable median. ( B ) Analysis of vaccinee sera by the cPass sVNT at time points t0–t5. The cut-off value according to the manufacturer’s manual is indicated by the dashed line (30% inhibition), and horizontal bars delineate the variable median. ( C ) Correlation of in-house sVNT and cPass sVNT values < 0% were set to 0%, and the correlation coefficient was calculated according to Spearman. Ih: in-house.

Article Snippet: Black high-binding 96-well plates (Greiner Bio-One GmbH, Frickenhausen, Germany) were coated with 200 ng/well of recombinant hACE2 (Sino Biological Inc., Beijing, China, #10108-H08H) diluted in 100 µL carbonate buffer at 4 °C for 16 h. The coated plates were washed four times using 300 µL/well TRIS-buffered saline (TBS)/0.1% Tween (TBS-T), blocked with 300 µL/well TBS-T/2% BSA ( w/v ) for 1.5 h at 37 °C, and washed four times using TBS-T.

Techniques: Construct, Binding Assay, Inhibition