ProteinChip® SELDI System

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ProteinChip® SELDI System


Question:

Why is normalization conducted on the ProteinChip® SELDI system? How is normalization done?


Answer:

Normalization is performed to minimize variation. The normalization process takes the total ion current used for all spots, averages the intensity, and adjusts the intensity scales for all spectra. Once the spectrum is normalized, the resulting normalization factor is used in downstream analysis, such as biomarker analysis.

The method of choice for normalization is total ion current (TIC), which uses the total area under the curve in a given range. The calculated normalization coefficient is the average TIC for all spectra. The calculated normalization factor is used to rescale the spectra so that all spectra have the same observed TIC.

Normalization to a reference peak is not recommended unless there is absolute certainty that the peak is at the same concentration in each and every sample.


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