Anti-respiratory syncytial virus (RSV) antibody cell-based potency assay
AssigneeMerck Sharp & Dohme Corp.
InventorsDai Wang, John P. Bilello, Kevin B. Gurney, Ping Han, Xi He, Dengyun Sun, Amy Hsu Tou
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Abstract
The disclosure describes methods for measuring the potency of anti-Respiratory Syncytial Virus (RSV) antibody against respiratory syncytial virus in a cell culture system using an RSV reporter virus. The disclosure also describes respiratory syncytial viruses that carry a reporter gene, and expression vectors for producing an infectious recombinant respiratory syncytial virus (RSV) that carry a reporter gene.
Claims21
An expression vector for producing an infectious recombinant respiratory syncytial virus (RSV) comprising:
- a) a nucleic acid sequence encoding a respiratory syncytial virus;
- and
- b) a reporter gene flanked by an RSV gene start sequence and an RSV gene end sequence, the reporter gene and flanking RSV gene start and RSV gene end sequences located between the P and M genes of the respiratory syncytial virus;
- wherein the RSV gene start sequence is SEQ ID NO:
- 16 and the RSV gene end sequence is SEQ ID NO:
- 17;
- and
- wherein the reporter gene encodes a luminescent enzyme that catalyzes a luminescent substrate;
- or the reporter gene encodes a fluorescent protein.
A respiratory syncytial virus comprising:
- a) a nucleic acid sequence encoding a respiratory syncytial virus;
- and
- b) a reporter gene flanked by an RSV gene start sequence and an RSV gene end sequence, the reporter gene and flanking RSV gene start and RSV gene end sequences located between the P and M genes of the respiratory syncytial virus;
- wherein the RSV gene start sequence is SEQ ID NO:
- 16 and the RSV gene end sequence is SEQ ID NO:
- 17;
- and
- wherein the reporter gene encodes a luminescent enzyme that catalyzes a luminescent substrate;
- or the reporter gene encodes a fluorescent protein.
A method for measuring the activity of an anti-respiratory syncytial virus (RSV) antibody or antigen binding fragment thereof, the method comprising:
- a) combining (i) the anti-RSV antibody or antigen binding fragment thereof, (ii) an RSV virus comprising a reporter gene, and (iii) one or more cells infectable by the RSV virus;
- and
- b) detecting expression of the reporter gene;
- wherein the RSV virus comprises
- i) a nucleic acid sequence encoding a respiratory syncytial virus;
- and
- ii) the reporter gene flanked by an RSV gene start sequence and an RSV gene end sequence, the reporter gene and flanking RSV gene start and RSV gene end sequences located between the P and M genes of the respiratory syncytial virus,
- wherein the reporter gene encodes a luminescent enzyme that catalyzes a luminescent substrate, and detecting expression of the reporter gene comprises detecting luminescence of the luminescent substrate, or wherein the reporter gene encodes a fluorescent protein, and detecting expression of the reporter gene comprises detecting fluorescent light emission from the fluorescent protein.
Dependencies are inferred from the claim text.