It is sometimes noticed that the virus sample gives peaks and sometimes not. The result of these two analyses sometimes may similar or a little bit modified. However, it supposed not to show any absorbance in virus sample. So how we can ensure by seeing the chromatogram after sample run which portion are containing maximum virus. Here, we used size exclusion and affinity based resin. Could anyone have experienced about virus sample run and its scenario in the chromatogram. Thanks in advance.
It seems very unlikely to me that virus particles would show no UV absorbance in a chromatogram, since virus contains a large amount of nucleic acid, which has UV absorbance peak at about 260 nm, and proteins, which have UV absorbance peak at about 280 nm. If you do not see an absorbance peak for virus, it could be due to a lack of sufficient virus in the sample to detect the peak, or to a high background absorbance from non-viral substances in the sample.
Ok with the annswers of B Shapiro;
Also, I think in the case or handling a live virus, try intracerebral inoculation of all samples collected to mice or suckling-mice depending on the rabie strain .
In case, you are using an inactivated virus , try ELISA or other suitable technique .
I am also ok with the answer of AB Shapiro. You can also check the work of Oita and Halewyck on Polio and the group of E Kenndler on Rhinovirus. They did CE but it should give you an idea what to expect when you run chromatography with UV detection...
You could try a Western blot of the different fractions as well. Obviously that won't give you real-time detection during the run but it might be helpful in troubleshooting
I agree that it would be very unlikely for a virus to have no UV absorbance. One can expect the UV spectra to have characteristic maxima at 260/280. However, keep in mind that you are not looking for the UV spectra, you are looking for UV response and that the UV response may be higher at 210 nm. Over the years I have done extensive work at 210 nm. It is possible that you will see more peaks at 210nm, and it may make your chromatography work more complex, but if you have a dilute sample, it may make the difference between having an assay and not having an assay. You might run your sample in a cuvette on a spectrophotometer to determine if this is feasible.
Thanks Ryan for your information. I have done some sample run according to your suggestion and I am happy that I have seen much higher response in 210nm in comparison to my previous wavelength setup . Hope after analysis I will get better result.
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