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The single immunization of chickens with the purified fusion proteins, at a dose equivalent to 10 μg of the knob moiety, elicited serum antibodies with high hemagglutination inhibition (HI) activities, similar to those induced by an inactivated EDS vaccine.
To confirm the reactivity patterns of AVFluIgG01 and AVFluIgG03, HI activities against H5N1, H3N2, and H1N1 viruses were assessed (Table 4).
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Based on the results of the HI assay, mAbs 2D9 and 4C2 were chosen for further studies due to their high HI activity (data not shown) against a wide range of rescued reassortant viruses from different clades.
It was found that addition of MgCl2 is an alternative approach to remove lysozyme from ovomucin, and that the hemagglutination inhibition (HI) activity of ovomucin with MgCl2 against New Disease Virus NDVV) was about two times higher than the protein without salts.
A dose response analysis indicated that a single immunization with as little as 1 μg of the knob moiety of the CMP knob fusion protein was as effective as the inactivated vaccine in inducing antibodies with HI activity.
We determined HI activity using turkey erythrocytes incubated with sera from vaccinated groups.
To do this, we tested the same sera against each of the three vaccine component strains of virus for hemagglutination-inhibitory (HI) activity (Fig. 3b).
Following this, at 2 months, we again see significant HI activity maintained in the Nanopatch™ vaccinated mice compared to the intramuscular injected groups.
The HI activity assay is the most widely accepted 'gold standard' used as the surrogate for influenza protective effectiveness [28], [29].
To further compare the systemic responses induced, we determined the HI activity of immunized mouse sera against both A/PR8 and A/Philippines viruses.
As shown in Fig. 5B, cVLPs containing the membrane-anchored flagellin also induced antibodies showing HI activity to A/Philippines virus (a mean value of 28).
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