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The Complement System pp 257–272Cite as

Purification of Native Surfactant Protein SP-A from Pooled Amniotic Fluid and Bronchoalveolar Lavage

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

Abstract

Surfactant protein SP-A is a hydrophilic glycoprotein, similar to SP-D, which plays an important role in pulmonary surfactant homeostasis and innate immunity. SP-A is actively expressed in the alveolar type II cells and Clara cells. Their basic structure consists of triple-helical collagen region and a C-terminal carbohydrate recognition domain (CRD). By binding to the infectious microbes, SP-A (like SP-D) are involved in pathogen opsonization and agglutination and subsequent clearance of the microorganism, via recruitment of phagocytic cells via receptors for the collagen region. SP-A has also been localized at extra-pulmonary sites such as salivary epithelium, amniotic fluid, prostate glands, and semen. The presence of SP-A in fetal and maternal tissue and amniotic fluid suggests it is involved in pregnancy and labor. Native SP-A can be purified from amniotic fluid and bronchiolar lavage fluid (BALF) via affinity chromatography. In addition, we also report here a procedure to express and purify a recombinant form of trimeric CRD in Escherichia coli. The availability of highly pure native SP-A and CRD region can be central to studies that examine the diverse roles that SP-A play in surfactant homeostasis, pulmonary infection and inflammation and pregnancy.

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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

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

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

    Priyaa Madhukaran & Taruna Madan

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

    Patrick Waters

Authors
  1. Najmunisa Karbani
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  2. Eswari Dodagatta-Marri
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  3. Asif S. Qaseem
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  4. Priyaa Madhukaran
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  5. Patrick Waters
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  6. Anthony G. Tsolaki
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  7. Taruna Madan
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  8. Uday Kishore
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Editor information

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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Karbani, N. et al. (2014). Purification of Native Surfactant Protein SP-A 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_21

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  • DOI: https://doi.org/10.1007/978-1-62703-724-2_21

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-723-5

  • Online ISBN: 978-1-62703-724-2

  • eBook Packages: Springer Protocols

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