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- Nachgewiesen in: USPTO Patent Applications
- Sprachen: English
- Document Number: 20230266320
- Publication Date: August 24, 2023
- Appl. No: 17/779011
- Application Filed: November 20, 2020
- Assignees: Merck Sharp & Dohme LLC (Rahway, NJ, US)
- Claim: 1. A method of detecting the presence of Cytomegalovirus (CMV) in a sample, comprising measuring the amount of pentameric complex in a sample using an indirect sandwich enzyme linked immunosorbent assay (ELISA), and quantifying the amount against a reference sample with a known amount of CMV, wherein the sample is a drug substance sample, drug product sample, or process intermediary sample.
- Claim: 2. The method of claim 1, wherein detecting and quantifying the presence of Cytomegalovirus (CMV) does not detect and quantify multiple glycoproteins.
- Claim: 3. The method of claim 1, wherein the pentameric complex comprises gH, gL, UL128, UL130, and UL131.
- Claim: 4. The method of claim 3, wherein the method additionally comprises P188.
- Claim: 5. The method of claim 1, wherein the sandwich ELISA comprises two monoclonal antibodies against different epitopes on the pentameric complex used to detect and quantitate the pentameric complex, wherein the two monoclonal antibodies comprise: a. a capture antibody, wherein the capture antibody is an IgG1 mouse-rabbit chimeric antibody, and b. a detection antibody selected from a human antibody isolated from patients with a naturally occurring CMV infection.
- Claim: 6. The method of claim 5, wherein the capture antibody is 57.4C.
- Claim: 7. The method of claim 5, wherein the detection antibody is selected from D1-103H or 25.2.
- Claim: 8. The method of claim 1, wherein the two antibodies are used to quantitate the glycoprotein complex comprising measuring the sample against relative potency of the sample is quantitated against the antigen content of a known reference standard.
- Claim: 9. A method for quantifying Cytomegalovirus comprising a sandwich ELISA assay measuring the amount of the pentameric complex, wherein the pentameric complex comprises gH, gL, UL128, UL130, and UL131, and wherein this amount is quantified relative to a reference standard of a sample with a known amount of CMV.
- Claim: 10. (canceled)
- Claim: 11. An assay for detecting Cytomegalovirus (CMV) in a sample comprising measuring and quantitating the glycoprotein complex comprising: a. coating an assay plate with an antigen b. adding a first monoclonal antibody and incubating, c. adding a reference standard and/or vaccine sample and incubating, d. washing the plate, e. adding a second monoclonal antibody, f. washing the plate, g. adding a fluorogenic substrate, h. calculating the glycoprotein complex, and i. quantitating the glycoprotein complex relative to a reference standard of a sample.
- Claim: 12. The assay of claim 11, wherein the first monoclonal antibody is 57.4C mouse-rabbit chimeric mAb.
- Claim: 13. The assay of claim 11, wherein the second monoclonal antibody is D1-103H human mAb or 25.2H human mAb.
- Claim: 14. The assay of claim 11, wherein the antigen is a polyclonal antibody.
- Claim: 15. The assay of claim 11, wherein the polyclonal antibody is a goat anti-mouse IgG1Fc polyclonal antibody.
- Claim: 16. The assay of claim 11, wherein the fluorogenic substrate is 4MUP.
- Claim: 17. A kit for detecting, measuring and quantitating the glycoprotein complex, wherein the kit comprises a. an ELISA plate, b. a goat anti-mouse IgGlFc polyclonal antibody, c. a detection antibody, wherein the detection antibody is a human antibody: d. a capture antibody, e. a fluorogenic substrate, f. an assay diluent g. 2X Balancing Buffer h. a storage buffer i. a coating buffer, and j. a wash buffer.
- Claim: 18. The kit according to claim 17, wherein the detection antibody is a human antibody selected from one of the following: D1-103H or 25.2 antibody and wherein the capture antibody is 57.4, an IgGl mouse-rabbit chimeric antibody.
- Claim: 19. (canceled)
- Claim: 20. The kit according to claim 17, wherein the fluorogenic substrate is 4MUP.
- Claim: 21. (canceled)
- Claim: 22. (canceled)
- Claim: 23. (canceled)
- Claim: 24. (canceled)
- Claim: 25. (canceled)
- Claim: 26. The kit according to claim 17, wherein a. the assay diluent comprises 10 mM Imidazole, 150 mM NaCl, 0.5% BSA, 0.05% P188 pH 7.0, b. the 2X balancing buffer comprises 20 mM Imidazole, 225 mM NaCl, 1% BSA, 0.1% P188, pH 7.0, c. the storage buffer comprises 25 mM Histidine, 75 mM NaCl, 9% w/v Sucrose, pH 7.0, d. the 2X balancing buffer comprises 20 mM Imidazole, 225 mM NaCl, 1% BSA, 0.1% P188, pH 7.0, e. the storage buffer comprises 25 mM Histidine, 75 mM NaCl, 9% w/v Sucrose, pH 7.0, f. the coating buffer comprises 1X TBS (25 mM Tris, 0.15 M NaCl, pH 7.2), and g. the wash buffer comprises 1X TBS (25 mM Tris, 0.15 M NaCl, pH 7.2).
- Current International Class: 01
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