Method for virus assay
The present invention relates to a method for virus assay. More closely the invention relates a method for total quantification of adenovirus in a sample as well as total and functional (active) adenovirus in a sample. The method for determining adenovirus concentration in a sample comprises subjecting said sample to SPR (surface plasmon resonance) assay with immobilized FX (Factor X) and/or immobilized CAR (coxsackievirus and adenovirus receptor) on a sensor surface, wherein the adenovirus concentration is determined from sample binding to immobilized FX and/or immobilized CAR. CAR can be replaced by an ligand binding to adenovirus fiber, such as an anti-adenovirus fiber antibody. FX can be replaced by a ligand binding to adenovirus hexon, such as an anti-adenovirus hexon antibody. The method can be used for quality control in an adenovirus purification process, for example for gene therapy.
1. A method for determining adenovirus concentration in a sample, comprising subjecting said sample to surface plasmon resonance (SPR) assay with immobilized Factor X (FX) and/or immobilized coxsackievirus and adenovirus receptor (CAR) on a sensor surface configured to determine the adenovirus concentration, wherein the adenovirus concentration is determined from sample binding to immobilized FX and/or immobilized CAR.
2. The method according to claim 1 , wherein FX or CAR are immobilized.
3. The method according to claim 1 , wherein said FX and said CAR are immobilized on different SPR surfaces or on the same SPR surface but on different regions or areas thereof.
4. The method according to claim 1 , wherein functional adenovirus is determined by binding to both FX and CAR by the same adenovirus.
5. The method according to claim 1 , wherein FX or CAR is immobilized and the one of FX and CAR that is not immobilized is injected in a second step of the SPR assay for binding to relevant parts of the sample (adenovirus) already bound to the surface in a sandwich assay.
6. The method according to claim 1 , wherein CAR is replaced by a ligand binding to adenovirus fiber.
7. The method according to claim 1 , wherein FX is replaced by a ligand binding to adenovirus hexon.
8. The method according to claim 1 , wherein the ratio between functional (active) adenovirus concentration and total adenovirus concentration is determined.
9. The method according to claim 1 , wherein the result from the SPR assay(s) are determined with standard curves.
10. The method according to claim 1 , wherein the SPR assay(s) is/are calibration free concentration analysis (CFCA) assays.
11. The method of claim 1 , wherein the adenovirus concentration is used for quality control in an adenovirus purification process.
12. The method of claim 1 , wherein the adenovirus concentration is used for quality control in a purification process of adenovirus for gene therapy.
13. The method according to claim 6 , wherein the ligand binding to adenovirus fiber is an anti-adenovirus fiber antibody.
14. The method according to claim 7 , wherein the ligand binding to adenovirus hexon is an anti-adenovirus hexon antibody.