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

1 Like
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

4

Are you also using the same plate washer? I would also compare for how long the reagents are standing on the Hamilton deck vs the time on the bench on the manual procedure. We have similar issue once and it was because, when performed manually, the reagent are use freshly. In the robot, the reagent say on the deck for a few our to achieve maximum throughput.

1 Like