We are facing a serious problem in which when ever we are performing a kinetic experiment by a capture method the capture level is drastically falling down from cycle to cycle.
Here we are immobilizing anti-Histidine antibody (50ug/100uL) on CM5 chip using specify contact time (420 Seconds) which results in 10 to 12 thousand Ru final response.
Capture used is FcRn.
IgG antibody as the analyte.
Regeneration is 10mM Glycine pH-1.5.
The run includes 6 dilutions in which four samples are run in one run i.e., in total 28 injections. The capture level is not remaining consistent from the first injection to the last injection.
Please refer the file attached for a better understanding.