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Purification of Surfactant Protein D (SP-D) from Pooled Amniotic Fluid and Bronchoalveolar Lavage

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The Complement System

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1100))

Abstract

Surfactant protein SP-D is a multimeric collagenous lectin, called collectin. SP-D is a multifunctional, pattern recognition innate immune molecule, which binds in a calcium dependent manner to an array of carbohydrates and lipids, thus offering resistance to invading pathogens, allergen challenge, and pulmonary inflammation. SP-D is predominantly found in the endoplasmic reticulum of type 2 pneumocytes and in the secretory granules of Clara or non-ciliated bronchiolar cells. The highest expression of SP-D is observed in the distal airways and alveoli. There is also an extra pulmonary existence of SP-D. The common sources of native full-length human SP-D are bronchoalveolar lavage (BAL) washings from normal or preferably patients suffering from alveolar proteinosis who overproduce SP-D in the lungs. Amniotic fluid collected at the term during parturition is another reasonable source. Here, we describe a simple and rapid method of purifying native SP-D away from SP-A which is also present in the same source. We also describe procedures of expressing and purifying a recombinant fragment of human SP-D (rhSP-D) comprising trimeric neck and carbohydrate recognition domains that has been shown to have therapeutic effects in murine models of allergy and infection.

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Authors and Affiliations

  1. Centre for Infection, Immunity and Disease Mechanisms, Biosciences, School of Health Sciences and Social Care, Brunel University, London, UK

    Eswari Dodagatta-Marri, Asif S. Qaseem, Najmunisa Karbani, Anthony G. Tsolaki & Uday Kishore

  2. Neuroimmunlogy Group, John Radcliffe Hospital, Oxford, UK

    Patrick Waters

  3. Innate Immunity Group, National Institute of Reproductive Health, Mumbai, India

    Taruna Madan

Authors
  1. Eswari Dodagatta-Marri
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  2. Asif S. Qaseem
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  3. Najmunisa Karbani
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  4. Anthony G. Tsolaki
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  5. Patrick Waters
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  6. Taruna Madan
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  7. Uday Kishore
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Editors and Affiliations

  1. Division of Infectious Diseases, Brigham and Women’s Hospital, Harvard University Harvard Medical School, Boston, Massachusetts, USA

    Mihaela Gadjeva

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Dodagatta-Marri, E. et al. (2014). Purification of Surfactant Protein D (SP-D) from Pooled Amniotic Fluid and Bronchoalveolar Lavage. In: Gadjeva, M. (eds) The Complement System. Methods in Molecular Biology, vol 1100. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-724-2_22

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  • DOI: https://doi.org/10.1007/978-1-62703-724-2_22

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-723-5

  • Online ISBN: 978-1-62703-724-2

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