Dear all,
i am interested in checking for the degradation of an anti-PDL antibody over time in biological fluid such as serum and urine. Are any protocols that you can suggest or share any thoughts on this? I am thinking of going for either ELISA or WESTERN BLOT (using an anti-Fc antibody for example in the case of WB), especially the first since i can quantify the protein more accurately. I am also thinking time points such as 30 min, 1h, 2h, 3h and 16-24 hours.
Thank you in advance for your thoughts and suggestions.
Best wishes,
Nikos
hope this link is useful
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3089497/
regards
Many thanks Hassan.
The issue is that i already have the protein in its neat format let's say and it will may be found in urine and blood sera. So i cannot perform a luciferase assay since i cannot tag the gene to it using a vector.
Any alternatives for having the protein already?
BW
Nikos
By degradation do you mean unfolding? Or something else? Like the half-life of a protein? Check out these papers for estimating protein half-life:
Article Determining Protein Half-Lives
Article Measuring In Vivo Protein Half-Life
A less sophisticated way would be to measure mRNA levels for that protein (e.g RT-PCR) and compare it to the protein levels present at a particular time-point (e.g Western Blot), with the appropriate controls (e.g loading, amount used, etc). That will give you some idea of the rate of degradation of the protein in an indirect manner.
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