What kind of column to use for “the affinity stripping” of a biomarker from urine. Anyone have procedures with culture medium from cancer cell lines producing biomarkers too?
Hi Emily, it depends on what biomarker you are after and the endpoint of this process.
The most selective way is to immobilize an antibody that specifically binds to your target.
There are other ways to enrich a sample for your biomarker, but that requires a description of what protein you are going for.
I'd like to remove the biomarker from urine so that I can see how much background signal contributes to the test signal in ELISA or lateral flow assays.
I already tried pH , immunoprecipitate, and heat treated the urine to get the biomarker to denature, but it doesn't look like it worked at all.
I am trying to remove he4.
You could try TCA/acetone precipitation of urine to remove proteins.
Article Evaluation of Urinary Protein Precipitation Protocols for th...
If you just want to see non-specific background, you could treat boiled urine samples with proteinase K/trypsin. Then inhibit them with PMSF and run the ELISA.
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