Hi, I am currently producing a recombinant protein from yeast Pichia pastoris. The medium used in the bioreactor is the Basal salt medium supplemented with PTM1 salts. I have noticed that when performing the ELISA test, my protein yield in the sample is quite low. I am guessing that the components in my medium are interfering with the analysis. Is it possible to perform a simple and inexpensive pre-processing step that can possibly remove the unwanted components from the sample so that the yield of my protein can be seen higher through the ELISA?
I'm not sure but if it can help you, https://www.nature.com/articles/srep28887
Suni Mathew thank you very much for forwarding the paper. I will have a look at it :)
Purification of proteins from Pichia may get tricky some times. Are you sure that the low intensity observed in ELISA is dues to interference? otherwise confirm the expression with a tag/protein specific Western blot. If salt is the main issue, your can try desalting columns and further concentrating of purified product after.
Regards,
Dhananjayan Dhanasooraj thank you very much for the reply. I am not 100% sure if the low intensity is due to the interference. However, from literature and discussions with people working within a similar area, it came down to such a possibility. At the moment I have only two options in the lab to check the proteins; (i) ELISA and (ii) HPLC. Do you recommend any specific column for the HPLC to quantify recombinant proteins produced in basal salts medium?
Hi Jewel Ann Joseph . I am not familiar with HPLC based quantification of proteins, sorry for the same. Hope you done SDS-PAGE and sure that no co-purified proteins are there.
Best regards,
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