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January 01, 2011 (Vol. 31, No. 1)

Automating Protein Sample Preparation

Perfinity Workstation Exploits Separation Technology to Streamline Critical Operation

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    Figure 1. Immunochromatographic analysis of transferrin in pooled human serum

    The world rejoiced at the completion of the Human Genome Project. Researchers are now faced with the challenges of the proteome, which is vastly more complex and data rich. The diagnostics industry is expanding upon traditional immunological assays, utilizing tests that discriminate isoforms and splice variants, while at the same time identifying post-translational modifications.

    In the pharmaceutical sciences, the industry-wide shift from small molecule therapies to biopharmaceuticals has resulted in drug therapies that are much more difficult to isolate with intricacies that are exceedingly difficult to detect. Mass spectrometers have the capability to resolve these complexities but only when preceded by separation processes that prepare proteins and peptides for analysis. A problem with current fractionation approaches is that they are complex, multifaceted, and often yield irreproducible results.

    Perfinity’s new Perfinity Workstation exploits novel separation technology to automate sample preparation. The Perfinity Workstation enables users to start with serum and have peptides ready for LC/MS analysis in as little as 10 minutes. This system dovetails nicely into most existing LC/MS workflows.

    The Perfinity Workstation consists of five columns. Each column performs a step of the mass-spec sample-preparation process—affinity selection, buffer exchange, digestion, desalting, and reverse-phase separation. Automated integration of these steps removes much of the equipment and labor associated with mass-spectral analyses of proteins (Figure 1).

    Standard immunoassays have been the dominant method of performing routine serum protein analyses for the last 60 years for good reason. They are a conceptually simple method capable of measuring antigens down to the pg/mL level.

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