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Evaluation of density gradient separation methods

W D Corry, P A Bresnahan, G V Seaman

    Journal of Biochemical and Biophysical Methods
    |December 1, 1982
    PubMed
    Summary
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    A novel inconsistency number (I) quantifies isopycnic separation effectiveness, with zero indicating perfect sorting. This method ranks red cell separation techniques, revealing performance variations based on operating conditions.

    Area of Science:

    • Biochemistry
    • Analytical Chemistry
    • Cell Biology

    Background:

    • Isopycnic separation is crucial for isolating biological components.
    • Evaluating the true effectiveness of these separation techniques remains challenging.
    • Existing methods may not accurately reflect the sorting capabilities of isopycnic processes.

    Purpose of the Study:

    • To develop and validate a new quantitative method for assessing isopycnic separation effectiveness.
    • To introduce an inconsistency number (I) as a performance metric.
    • To compare the performance of different red blood cell separation techniques.

    Main Methods:

    • A novel inconsistency number (I) was developed, ranging from 0 (perfect separation) to 1 (sample subdivision).
    • The method's validity was confirmed by observing performance changes in a known separation technique.

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  • Four distinct red blood cell separation methods were evaluated using the inconsistency number.

    • Main Results:

    • The inconsistency number (I) effectively reflects changes in separation performance.
    • A performance hierarchy for red cell ordering by density was established: Murphy technique (I=0.21) > albumin density gradient (I=0.43) > Stractan density gradient (I=0.53) > Percoll density gradient (I=0.73).
    • Separation performance was sensitive to specific operating conditions and the isolated red cell fraction.

    Conclusions:

    • The inconsistency number (I) provides a reliable metric for evaluating isopycnic separation processes.
    • The study establishes a clear performance ranking of common red blood cell separation techniques.
    • Optimization of operating conditions is critical for maximizing the efficiency of density-based cell separation.