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Purification, Quantification, and Functional Analysis of Collectins

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Lectin Purification and Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2132))

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Abstract

Native and recombinant collectins are purified by using mannan-agarose and an anti-collectin antibody column. The use of sandwich enzyme-linked immunosorbent assay (ELISA) with two antibodies against human mannan-binding lectin (MBL) enables elucidation of the collectin concentration in the blood, serum, and plasma. The collectin sugar specificity is demonstrated by determining the concentration of saccharide required to inhibit sugar binding by 50% in a saccharide-binding assay. Biological analyses including the complement-dependent hemolysis test and several other methods are used to evaluate collectin.

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Acknowledgments

This work was supported by grants from JSPS KAKENHI (to K.O.: grant numbers 10020801, 26460353, 17 K08615; to N.W.: grant numbers 22390113, 26293124, 17H04108), the Smoking Research Foundation, and the Mizutani Foundation for Glycoscience.

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Authors and Affiliations

  1. Department of Clinical Nutrition, Rakuno Gakuen University, Hokkaido, Japan

    Katsuki Ohtani

  2. Department of Medicine and Physiology, Rakuno Gakuen University, Hokkaido, Japan

    Nobutaka Wakamiya

Authors
  1. Katsuki Ohtani
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  2. Nobutaka Wakamiya
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Corresponding author

Correspondence to Katsuki Ohtani .

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Editors and Affiliations

  1. Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan

    Jun Hirabayashi

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Ohtani, K., Wakamiya, N. (2020). Purification, Quantification, and Functional Analysis of Collectins. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_10

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  • DOI: https://doi.org/10.1007/978-1-0716-0430-4_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0429-8

  • Online ISBN: 978-1-0716-0430-4

  • eBook Packages: Springer Protocols

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