I am working on a development of blood-based biomarker protein for AD and ALS; however, protein candidate is not readily available in purified form. Therefore, I have the problem of not having calibration curve to quantitate this protein. I was thinking that if I collect , say 50 plasma samples and I know that the bio-marker protein is elevated. Pooled these plasma and construct a calibration curve based on their total protein content. Now, I have a kind of dose/response curve. I am asking my fellow researchers that is this way a legit. I know it is not ideal, but i stacked with the absence of the purified form of this protein. We have attempted, but it is quite unstable. Any recommendation, thoughts, tips are welcome.
Best
Baki
Hello Agbas,
In principle, yes, you can try this way. But you need to determine the dilutional recovery of these pooled plasma (undilution, 2x, 4x, 8x, etc) in the first place. For instance, you see nice recovery after 2xdilution, then 2xdiluted pooled plasam can be used as the first standard point of the curve. You may assign it as 1000ng/ml, 1000ug/ml or whatever you like to quantitate your target protein.
However, I would recommend you synthesize the peptide recognized by the antibody in your assay if you by chance know the epitope. Otherwise, just try the way abovementioned.
I hope it helps.
Roy
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