التفاصيل البيبلوغرافية
| العنوان: |
ASSAYS FOR INFLUENZA VIRUS HEMAGGLUTININS |
| Document Number: |
20120237545 |
| تاريخ النشر: |
September 20, 2012 |
| Appl. No: |
13/322913 |
| Application Filed: |
May 28, 2010 |
| مستخلص: |
A combination of ultrafiltration and HPLC is used to analyze influenza virus. This combination is able to quantify hemagglutinin (HA) and correlates well with single radial immunodiffusion (SRID) results, but can be performed without the delay of waiting for immunochemical SRID reagents. |
| Inventors: |
Dormitzer, Philip (Weston, MA, US); Wen, Yingxia (Acton, MA, US); Rinella, Paola (Figline Valdarno, IT); Palmer, Gene (Sommerville, MA, US) |
| Assignees: |
NORVARTIS AG (Basel, CH) |
| Claim: |
1. A method for purifying influenza virus HA in a sample, comprising steps of: (i) ultrafiltration of the sample to provide a filtrate; and (ii) RP-HPLC of the filtrate to separate any HA therein from any other components therein. |
| Claim: |
2. The method of claim 1, wherein step (i) uses an ultrafiltration membrane with a 300 kDa cut-off. |
| Claim: |
3. The method of claim 1, wherein step (ii) uses a RP-HPLC support of polystyrenedivinylbenzene particles with a pore size between 500-5000 Å. |
| Claim: |
4. The method of claim 1, wherein the sample contains HA from only one influenza virus strain. |
| Claim: |
5. The method of claim 1, wherein the HA comprises influenza A virus HA. |
| Claim: |
6. The method of claim 5, wherein the HA is H1, H3 or H5 HA. |
| Claim: |
7. The method of claim 1, wherein the HA comprises influenza B virus HA. |
| Claim: |
8. The method of claim 1, wherein HA in the sample is from an influenza virion. |
| Claim: |
9. The method of claim 1, wherein the sample includes (i) influenza virus HA and (ii) influenza virus antigens PB1, PB2, PA, NP, NA, M1, M2, NS1 and/or NS2. |
| Claim: |
10. The method of claim 1, wherein the sample includes whole influenza virions. |
| Claim: |
11. The method of claim 1, wherein the sample includes split influenza virions. |
| Claim: |
12. The method of claim 1, wherein the sample includes purified surface antigens from influenza virions. |
| Claim: |
13. The method of claim 1, wherein the HA is HA1. |
| Claim: |
14. The method of claim 1, wherein the sample is free from (i) serum components, (ii) egg proteins, and/or (iii) chicken DNA. |
| Claim: |
15. The method of claim 1, wherein the sample includes no viral proteins except for influenza virus proteins. |
| Claim: |
16. The method of claim 1, wherein the sample includes detergent. |
| Claim: |
17. The method of any 1, wherein the sample is a bulk vaccine. |
| Claim: |
18. A method for analyzing a vaccine comprising steps of: (a) purifying HA in the vaccine, or in a sample thereof, by the method of claim 1; and (b) using the results of step (a) to calculate the HA concentration in the vaccine. |
| Claim: |
19. A method for providing a bulk vaccine with a desired HA concentration, comprising steps of: (a) purifying HA in the vaccine, or in a sample thereof, by the method of claim 16; (b) using the results of step (a) to calculate the HA concentration in the bulk vaccine; and (c) using the results of step (b) to dilute the bulk vaccine to give the desired HA concentration. |
| Claim: |
20. The method of claim 19, wherein the desired HA concentration is between 1 and 150 μg/ml e.g. 90 μg/ml, 45 μg/ml, 30 μg/ml, 15 μg/ml, 10 μg/ml, 7.5 μg/ml, 5 μg/ml, 3.8 μg/ml, 3.75 μg/ml, 1.9 μg/ml, or 1.5 μg/ml. |
| Claim: |
21. A method for providing a vaccine for patient use, comprising steps of: preparing a bulk vaccine with a desired HA content by the method of claim 19; and then extracting one or more unit doses of vaccine from the diluted bulk. |
| Claim: |
22. The method of claim 21, wherein an extracted unit dose is packaged as a kit component in combination with a second kit component, wherein the second kit component is a vaccine adjuvant. |
| Claim: |
23. A method for providing a bulk adjuvanted vaccine comprising steps of: preparing a bulk vaccine with a desired HA content by the method of claim 19; and then mixing the diluted bulk vaccine with an adjuvant. |
| Claim: |
24. A method for providing an adjuvanted vaccine for patient use, comprising steps of: providing a bulk adjuvanted vaccine by the method of claim 23; and then extracting one or more unit doses of vaccine from the bulk. |
| Current U.S. Class: |
4242/101 |
| Current International Class: |
61; 61; 61; 12; 12 |
| رقم الانضمام: |
edspap.20120237545 |
| قاعدة البيانات: |
USPTO Patent Applications |