MSD Assay Signal Lost

1

Hello All,

I am just wondering if anyone here automated an MSD assay. The problem I am running is that, when we run the assay on Hamilton the signal lost by 4-5 times compare to manual run., we are using same samples and same plate reader. I checked and all pipetting is correct and accurate and cv’s are less than 10% and incubation time and temperature is correct.

I am just wondering if anyone have run in this problem and have any thoughts or suggestions for me to try.

Thanks

2

I am not an MSD assay expert, but I just finished supporting the automation of one. I had similar issues in early development run with controls returning low signal. We found that while the liquid handler was accurate, the manual pipetting was likely over delivering sample. I would recommend comparing the volumes on of the analyst with a hand pipette of sample matrix vs the Hamilton.

We wound up having to adjust the liquid class to match the hand pipette, even if it was less accurate. This was a compromise to have the automated data match the validated manual results.

3

Thanks a lot for the information sean, this is very intersting and a little disappointing but will look into it