I have made TL reagent by mixing antibody in 1X PBS containing 0.25% sucrose for striping similarly the CL which is anti species of TL and prepared using the above composition. While running the LFA, I can see both TL and CL increases linearly with my analyte concentration which was strange.

I have found out with my investigation that my test line antibody move during the running of the lateral flow and the TL antibody gets captured at the CL and forms the sandwich with the analyte there and hence I see the linear correlation with the analyte.

I need to know how can I immobilize my antibody firmly to the nitrocellulose membrane, such that it does not move and interfere with CL antibody.

Thanks,

Yogita