Solid liquid liquid extraction of porcine gastric mucins from homogenized animal material

Abstract

Mucins purified from porcine stomachs have gained importance in biomedical applications as they exhibit unique features such as hydrogel formation, lubricity and antivirality. Because commercially available porcine gastric mucins (PGM) are neither gel forming, nor do they reduce friction, a robust purification process for functional mucins from porcine tissue (mainly Muc5AC) is necessary. Based on our former investigations (V. J. Schömig, B. T. Käsdorf, C. Scholz, K. Bidmon, O. Lieleg and S. Berensmeier, RSC Adv., 2016, 6, 44932-44943), we further optimized the established purification process in terms of productivity and overall yield as well as ease of scalability. We therefore introduced a novel extraction method -solid liquid liquid extraction (SLLE) -as an early capture step from homogenized porcine stomach, which combines conventional solid liquid extraction with a second, immiscible solvent, to simultaneously delipidate the tissue and extract hydrophilic proteins into the polar phase. Using Design of Experiments (DoE) the parameters incubation time and ratio of solvent phases (hexane/water) were identified to 3 h and 1/15 for SLLE, respectively. PGM was collected in the polar phase and further purified by size exclusion chromatography and diafiltration. With the homogenization of porcine stomach and the introduction of the SLLE, up to 3570 mg mucin per stomach can be purified, resulting in 55 times more PGM compared to the former published reference process. The productivity of the process was up to 25 mg mucin per stomach and per h, representing an improved process productivity by a factor of 4. Lubricity and gel formation of the purified mucin were retained with the optimized extraction protocol.

Reversible Condensation of Mucins into Nanoparticles

“…In the compaction model, we hypothesize that the strong compaction observed is driven by poor solubility of mucins in glycerol. Indeed, mucins are typically insoluble in poorly polar solvents such as hexane and simple sugars, which are essential for the solubility of the mucin protein, , and are also less soluble in glycerol than in water . Increasing the glycerol concentration would then favor mucin intramolecular interactions over mucin–solvent interactions, leading to a compaction of the molecular structure.…”

Mucins as multifunctional building blocks of biomaterials

“…26 They can be extracted from mucosal tissues either by collecting the mucus itself and extracting the mucins, 27 or by homogenizing whole tissues. [28][29][30] Purification is achieved by leveraging mucins' unique physical-chemical characteristics, including their solubility profiles in solvents, large size, and strong net negative charge. Although the glycosylated fractions of mucins are relatively resistant to proteolytic degradation, the unglycosylated sections are more easily degraded, 31 which can alter the gelling properties of the mucins.…”

A Pandemic Early Warning System Decision Analysis Concept Utilizing a Distributed Network of Air Samplers via Electrostatic Air Precipitation

“…One such potential solvent would be a 50:50 water-hexane mixture. Solid-liquid separation of porcine gastric mucins was successfully demonstrated to extract proteins [16]. However, the use of polar and non-polar substances together requires another trick discussed later, known as coaxial electrospray.…”