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Related Experiment Video

Updated: Dec 23, 2025

A Quantitative Glycomics and Proteomics Combined Purification Strategy
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Purification, Quantification, and Functional Analysis of Collectins.

Katsuki Ohtani1, Nobutaka Wakamiya2

  • 1Department of Clinical Nutrition, Rakuno Gakuen University, Hokkaido, Japan. ohtani@rakuno.ac.jp.

Methods in Molecular Biology (Clifton, N.J.)
|April 20, 2020
PubMed
Summary

This study details methods for purifying and quantifying collectins, including human mannan-binding lectin (MBL). These techniques allow for the assessment of collectin concentration and sugar-binding specificity in biological samples.

Keywords:
C-type lectinCL-K1CL-L1CollectinComplementELISAInfluenza A virusMBL

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Last Updated: Dec 23, 2025

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Area of Science:

  • Biochemistry
  • Immunology
  • Molecular Biology

Background:

  • Collectins are a family of C-type lectins involved in innate immunity.
  • Accurate quantification and characterization of collectins are crucial for understanding their biological roles.

Purpose of the Study:

  • To establish robust methods for the purification and quantification of native and recombinant collectins.
  • To characterize the sugar-binding specificity and biological activity of collectins.

Main Methods:

  • Purification using mannan-agarose affinity chromatography and anti-collectin antibody columns.
  • Quantification via sandwich enzyme-linked immunosorbent assay (ELISA) for human mannan-binding lectin (MBL).
  • Saccharide-binding assays to determine 50% inhibitory concentrations.
  • Biological activity assessment using complement-dependent hemolysis tests.

Main Results:

  • Successful purification of native and recombinant collectins.
  • Development of a sensitive ELISA for MBL quantification in blood, serum, and plasma.
  • Characterization of collectin sugar specificity and functional activity.

Conclusions:

  • The described methods provide a comprehensive approach for collectin analysis.
  • These techniques are valuable for research in innate immunity and related clinical applications.