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neutralizing antibodies assay design


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#1 emmanuelle

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Posted 08 February 2011 - 10:50 AM

hi,
i'm gonna do such a neutralizing antibodies assay. But it's hard to decide the dilution of serum and virus sup. nobody in our lab did this before. Any one has an idea that how to choose the dilution and volume of the serum? and also the virus supernatant?

Thank you very much!

#2 BioMiha

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Posted 08 February 2011 - 11:30 PM

First do a titration curve of the virus supernatant. Then choose a virus titer that gives you roughly half the maximal value and use this concentration to do a titration curve of the serum.

#3 Jordan W

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Posted 10 February 2011 - 04:13 PM

First do a titration curve of the virus supernatant. Then choose a virus titer that gives you roughly half the maximal value and use this concentration to do a titration curve of the serum.



This titration curve is often referred to as a TCID50. Start with putting straight virus on your cells, dilute 5 fold in each well. There should be some dilution that gives half of maximal infectivity. In either PFU, or some other metric.

As for your antibody serum, you might want to test whether the serum alone (in this case, probably the uninnoculated mouse) gives you any neutralization. If not, then you can just add pure serum to the virus and see if you get any neutralization.




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