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Second-generation nanofiltered plasma-derived mannan-binding lectin product: process and characteristics.

I Laursen1, G Houen, P Højrup

  • 1Statens Serum Institut, Copenhagen, Denmark. inl@ssi.dk

Vox Sanguinis
|April 26, 2007
PubMed
Summary

A new second-generation Mannan-binding lectin (MBL) product was developed using improved purification and nanofiltration methods. This enhanced MBL therapy offers increased safety for patients with MBL deficiency and recurrent infections.

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Area of Science:

  • Immunology
  • Biochemistry
  • Protein Chemistry

Background:

  • Mannan-binding lectin (MBL) is crucial for innate immunity, aiding pathogen recognition and complement activation.
  • Low MBL levels increase susceptibility to severe infections, making MBL substitution therapy a promising treatment.
  • A first-generation MBL product demonstrated safety and patient benefit.

Purpose of the Study:

  • To develop a nanofiltered second-generation MBL product with enhanced safety and efficacy.
  • To optimize MBL purification using a novel affinity matrix.
  • To characterize the improved MBL product.

Main Methods:

  • Screening of carbohydrate-based gels for affinity matrices.
  • MBL extraction from fraction III and purification using Superdex 200 pg.

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  • Two virus reduction steps: Planova 20N nanofiltration and solvent/detergent treatment.
  • Further purification by anion-exchange and gel-filtration chromatography.

    • Main Results:

    • Superdex 200 pg proved superior for MBL affinity purification from fraction III, achieving a 70% yield.
    • Nanofiltration through Planova 20N significantly increased viral safety with minimal MBL loss.
    • The final MBL fraction (53% purity) contained high-oligomeric MBL, associated proteases (MASP), and a minor protein impurity (alpha2-macroglobulin).
    • Functional activity (mannan-binding, opsonic) remained intact, but C4 activation capacity was reduced by half.

    Conclusions:

    • A second-generation MBL production process was successfully developed, yielding an average of 50%.
    • Nanofiltration of MBL-MASP complexes through Planova 20N enhanced product safety with minimal MBL loss.