A Detection and Quantification Label-free Tool to Speed Up Downstream Processing of Model Mucins

Carvalho SB, et al., 13(1):e0190974, PLoS One, 2018

Mucins are high-molecular weight glycoproteins secreted by epithelial cells. Recent studies have linked mucins to a variety of inflammatory diseases and cancer, suggesting them to be good therapeutic targets. Described herein is the development of an efficient Bio-Layer Interferometry (BLI) assay to detect and quantify mucin, in order to speed up downstream processing. Two distinct mucins, MUC5B human Mucin and Bovine Submaxillary gland Mucin (BSM) were assessed in this study. Assays were designed using the lectin-mucin binding interaction. Lectins are sugar-binding proteins that can be used to detect carbohydrate structures. A Pall ForteBio Octet RED96 platform equipped with High Precision Streptavidin (SAX) Biosensor probes were used to perform BLI experiments. In order to select the most suitable lectin for mucin detection, a series of biotinylated lectins was immobilized onto SAX sensor tips. Subsequently, biotinylated lectin loaded sensor tips were dipped in mucin samples (BSM and MUC5B) and association-dissociation profiles were recorded. BLI data obtained were fitted to a 1:1 binding model. Overall results of this study suggest that this BLI technology based strategy can be applied for mucin purification strategies in downstream process development.

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