Alto Nicoya

Hello dears! Does anyone have an Alto Nicoya? Are you getting beautiful sensorgrams and data? This is a very interesting question for me. Sometimes I can see not so bad data but I can't get reproducibility, linearity, accuracy.
Thank You

 

Hi,
Can you elaborate a bit more on what you are measuring? Please show one or two sensorgrams? A good and a bad. How do you measure linearity and accuracy (and what is the purpose)?

Kind regards
Arnoud

Hello! My ligand is a cytokine and my analyte is a MAb. I can only guess why the same interaction is always measured differently. As for linearity and accuracy, I see this by indirect signs in order to understand the validity of the device.
Thank u so much

OK, it is a start….
The first sensorgram looks ok to me. Five concentrations of mab (concentrations: ? nM) and a slow dissociation. The sensorgram could improve by a longer association time for each analyte concentration to get more curvature in the traces. As for the second figure (first time I see sensorgrams of the Nicoya Alto) has very different traces. Most seem to suffer from drift or jumps and dilutions problems.
With these tiny amounts of reagents used in this system you must make sure that everything is a clean as possible. I recommend to spin the mab at least 5’ at 16000 xg to spin down possible aggregates. To minimize the drift, I would recommend a longer pre-wash before starting the analyte incubations.
Please share some more information about the second figure (what do we see, did you try, concentrations, injection times, regeneration etc.) 
Kind regards
Arnoud

Interestingly, some of this sensorograms contain the same ligand, analyte and solutions. But look very different. Alto produces 5 triple dilutions automatically. From one sample, for example, 1000 nM. Thus, the jumps between dilutions defy explanation. As a result, now Alto doesn't work.
Thanks for your reply! We can talk about SPR outside of Alto