heat inactivation could denature your proteins and affect binding by ELISA. However, it depends on what you are testing for in your ELISA. As for antibodies, we inactivate our serum samples for 30min @56C which works well and does not affect antibody binding in our ELISAs. Several groups have looked at the effect of heat inactivation on influenza viruses. Zou et al. Virol. J. 10:289 (2013), have shown that heat treating avian influenza viruses (H7N9) @56C for 30min or UV irradiation for 30min resulted in complete loss of infectivity. You can treat your supernatant with heat or UV and then look for infectious virus to make sure that you have inactivated the virus prior to running in your ELISA. As a control, you can spike a positive sample in culture supernatant, treat with various heat or UV conditions, and compare to untreated to see if this effects your ELISA.

Hi Alex,

I agree that heat inactivation would denature your proteins (the point of heat inactivation), which may then make them difficult to test by ELISA. The ELISA antibody pair you use would be the finally judge of that, but Abs that bind to denatured proteins are typically different than ones that bind well to the native proteins.

Have you tried inactivating your influenza virus using UV? That works wonderfully, and leaves the proteins in your solution intact for ELISA. This is often done in virus studies. UV cross-links the virus genomic RNA and renders it useless, but doesn't affect proteins. I'm happy to send a protocol about how to do this.

heat inactivation could denature your proteins and affect binding by ELISA. However, it depends on what you are testing for in your ELISA. As for antibodies, we inactivate our serum samples for 30min @56C which works well and does not affect antibody binding in our ELISAs. Several groups have looked at the effect of heat inactivation on influenza viruses. Zou et al. Virol. J. 10:289 (2013), have shown that heat treating avian influenza viruses (H7N9) @56C for 30min or UV irradiation for 30min resulted in complete loss of infectivity. You can treat your supernatant with heat or UV and then look for infectious virus to make sure that you have inactivated the virus prior to running in your ELISA. As a control, you can spike a positive sample in culture supernatant, treat with various heat or UV conditions, and compare to untreated to see if this effects your ELISA.