Recombinant monoclonal antibody specifically binds to IAV HA, expressed in Chinese Hamster Ovary cells(CHO).
Figure 1 Low pH induced conformational changes of H3 rHA were inhibited by the human serum samples.
Inhibition of low pH induced H3 rHA conformation change by a convenient human serum pool (Pool) in HCCIA. H3 rHAs bound to nickel-coated plates were treated with 100 μl of 200 ng/ml trypsin to cleave HA0 into HA1 and HA2. rHA coated plates were incubated with diluent only, 1:4000, 1:400, or 1:40 diluted Pool for 1 hour. The plate was washed and treated with a range of pH buffers followed by fixation with 0.05% glutaraldehyde/PBS. An ELISA was performed using a pH-specific mAb, HC31, and detected by measuring the OD at 450 nm.
Trost, J. F., LeMasters, E. H., Liu, F., Carney, P., Lu, X., Sugawara, K., ... & Katz, J. M. (2018). Development of a high-throughput assay to detect antibody inhibition of low pH induced conformational changes of influenza virus hemagglutinin. PloS one, 13(6), e0199683.
Figure 2 CCI showed virus neutralizing activity in TMN after removal of head-binding antibodies.
Correct folding of major epitopes for GH HA1 from A/Hong Kong/1/68 (H3N2) was confirmed by appropriate Ab binding profiles. GH HA1 from A/Hong Kong/1/68 (H3N2) and ectodomain H3 rHA from A/Aichi/1/68 (H3N2) were coated on a nickel-coated plate, an ELISA was performed by using rabbit antisera and a panel of conformation specific mAbs.
Trost, J. F., LeMasters, E. H., Liu, F., Carney, P., Lu, X., Sugawara, K., ... & Katz, J. M. (2018). Development of a high-throughput assay to detect antibody inhibition of low pH induced conformational changes of influenza virus hemagglutinin. PloS one, 13(6), e0199683.
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