Isolation, Physicochemical Characterization, Labeling, and Biological Evaluation of Mannans and Glucans

Michael D Kruppa; Douglas W Lowman; Harry E Ensley; Zuchao Ma; Bridget Graves; Jennifer Kintner; Jennifer V Hall; Tammy R Ozment; David L Williams

doi:10.1007/978-1-0716-2549-1_24

Isolation, Physicochemical Characterization, Labeling, and Biological Evaluation of Mannans and Glucans

Methods Mol Biol. 2022:2542:323-360. doi: 10.1007/978-1-0716-2549-1_24.

Authors

Michael D Kruppa¹, Douglas W Lowman², Harry E Ensley², Zuchao Ma², Bridget Graves³, Jennifer Kintner⁴, Jennifer V Hall⁵, Tammy R Ozment², David L Williams²

Affiliations

¹ Department of Biomedical Sciences, Center of Excellence in Inflammation, Infectious Disease and Immunity, Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA. Kruppa@etsu.edu.
² Department of Surgery, Center of Excellence in Inflammation, Infectious Disease and Immunity, Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.
³ Department of Surgery, Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.
⁴ Department of Biomedical Sciences, Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.
⁵ Department of Biomedical Sciences, Center of Excellence in Inflammation, Infectious Disease and Immunity, Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.

PMID: 36008676
DOI: 10.1007/978-1-0716-2549-1_24

Abstract

The cell wall contains mannans and glucans that are recognized by the host immune system. In this chapter, we will describe the methods to isolate mannans and glucans from the C. albicans cell wall. In addition, we describe how to determine purity, molecular size, and structure of the mannans and glucans. We also detail how to prepare the carbohydrates for in vitro, ex vivo, or in vivo use by describing endotoxin removal (depyrogenation), derivatization, and labeling and evaluation of bioactivity.

Keywords: Bio-Layer Interferometry; Dectin-1; Dectin-2; Gel permeation chromatography; Glucan; Mannan; Mannose receptor; NMR.

MeSH terms

Candida albicans
Cell Wall / chemistry
Glucans* / analysis
Mannans*

Substances

Glucans
Mannans