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Method for continuous purification of biological material using immunosorbent.

J J Hughes, S E Charm

    Biotechnology and Bioengineering
    |August 1, 1979
    PubMed
    Summary
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    A new continuous purification device uses antibody-bound agarose in pouches on a Mylar belt. This system efficiently isolates biological materials but faces challenges with binding capacity due to pouch diffusion and agarose resuspension.

    Area of Science:

    • Biotechnology
    • Biochemical Engineering
    • Immunochemistry

    Background:

    • Continuous purification methods are crucial for efficient biological material processing.
    • Immunosorbent-based purification offers high specificity for target molecules.
    • Existing batch methods can be time-consuming and labor-intensive.

    Purpose of the Study:

    • To develop and evaluate a novel mechanical device for continuous immunosorbent-based purification.
    • To adapt an existing batch purification protocol for human placental alkaline phosphatase to a continuous system.
    • To identify and address limitations in the continuous purification process.

    Main Methods:

    • A continuous purification system was designed using heat-sealed nylon pouches with agarose-bound antibody on a Mylar belt.

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  • The device featured a four-chamber sequential system for binding, dissociation, and regeneration.
  • An existing batch method for human placental alkaline phosphatase purification was adapted for the continuous device.

    • Main Results:

    • The continuous system demonstrated antigen binding and dissociation capabilities.
    • A decrease in antigen binding capacity was observed compared to free immunosorbents, attributed to pouch diffusion resistance and short exposure times.
    • Reduced binding capacity between successive belt passes resulted from incomplete agarose resuspension due to inefficient agitation and pouch squeezing.

    Conclusions:

    • The developed device offers a continuous approach to immunosorbent purification.
    • Pouch design and process parameters significantly impact binding efficiency.
    • Further optimization is needed to overcome limitations in diffusion, exposure time, and agarose resuspension for improved performance.