I'm using commercial Herceptin for some experiments and when running FPLC (column superdex 200, PBS mobile phase, eluting 0.5ml/min) on my samples I find, besides the antibody peak (UV 280nm) at 26-27 minutes, a peak at around 36-37 minutes (corresponding to a mass around 10 000, i think). Anyone else who have had the same problem and might know what this contamination could be?
It will depend on whose Herceptin preparation you are using. Some contain Chinese Hamster Ovary (CHO) cell proteins. Look in the contraindication info as it may tell you what other components are present if it is not CHO protein(s)
And if you want it pure, you could take fractions discarding the contaminant and check if it's affecting your experiments.
Dear Professor Aneheim
We bought Herceptin (a vial of powder for concentrate for solution for infusion) from Roche Ltd. (Switzerland). We dialyzed it against PBS buffer and we did not observe any contamination in FPLC. I think the opinion of Dr. Worrall is correct and in this case you can use the method described by Dr. Jover or you can buy Herceptin from Roche Company like us.
For anyone interested it was actually so that the buffer used for diluting the samle gave a large peak if it was not the same as the mobile phase. This peak showed up much earlier in the chromatogram than what could be expected for such small molecules (citrate and/or acetate in our case) which is why we mistakenly thought it was some sort of contaminant or similar. Anyway, switching sample buffer or mobile phase so they match solved the issue!
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