Immobilization of sugars in supermacroporous cryogels for the purification of lectins by affinity chromatography

Gonçalves, Gabriel Ramos Ferreira; Gandolfi, Olga Reinert Ramos; Santos, Leandro Soares; Bonomo, Renata Cristina Ferreira; Veloso, Cristiane Martins; Veríssimo, Lizzy Ayra Alcântara; Fontan, Rafael da Costa Ilhéu

doi:10.1016/j.jchromb.2017.10.019

Cited by 23 publications

(7 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition to GC, HPLC and UPLC methods, affinity chromatography (AC) was also used, which is a type of liquid chromatography based on the use of specific chemical reactions between chemically reactive ligands bond to the stationary phase and atypical components of the mobile phase. It is generally used to separate proteins, e.g., lectins (Gonçalves et al 2017;Perret and Boschetti 2018). The lectins were isolated, among others from R. pseudoacacia seeds with the use of AC with ovomucin and elution with different buffers (Fleischmann and Rudiger 1986).…”

Section: Black Locustmentioning

confidence: 99%

Characterization of bioactive compounds in the biomass of black locust, poplar and willow

Tyśkiewicz

Konkol

Kowalski

et al. 2019

Trees

View full text Add to dashboard Cite

Key message Liquid chromatography is the main method for assaying bioactive compounds in biomass. Total phenolic content and antioxidant capacity are measured with spectroscopy. Phenolics are the most important bioactive substances in short rotation coppice. Secondary metabolites of black locust, willow and poplar have many different activities. The richest source of bioactive compounds are flowers, bark and buds. Abstract The black locust (Robinia pseudoacacia), willow (Salix spp.) and poplar (Populus spp.) may be the source of the biomass obtained from natural habitats as well as intentionally established plantations in the system of short rotation coppice. These species are promising candidates as an alternative source of bioactive compounds in researches for developing new bioproducts for multidirectional uses, including polymers, lubricants, construction materials, pharmaceuticals, as well as bioenergy and fuels. Various parts of plants, such as leaves, flowers, seeds, bark, woods, buds, catkins, whole tree material and bee products (honey, propolis) are a source of bioactive compounds. The characterization of these compounds, especially phenolic compounds (flavonoids, stilbenes, lignans, phenolic acids, tannins and phenolic glucosides), fatty acids, sterols, etc. may be determined by a variety of analytical (spectroscopy, chromatography) and extraction methods. This review contains 131 references and systematize all available data on the characterization of the bioactive compounds in the biomass of black locust, poplar and willow by analytical and extraction methods.

show abstract

Section: Black Locustmentioning

confidence: 99%

Characterization of bioactive compounds in the biomass of black locust, poplar and willow

Tyśkiewicz

Konkol

Kowalski

et al. 2019

Trees

View full text Add to dashboard Cite

show abstract

“…Subsequently, they are washed for the removal of unbound or weakly bound compounds and then passed to the mobile phase under desorption conditions, reversing the specific bond that had been formed. Because this is a widely used technique for the purification of lectins which bind to carbohydrates or glycoproteins coupled in the matrix, according to specificity, these molecules are eaten with carbohydrates or glycoproteins in a solution; for example, the Sephadex ® G-50 matrix retains lectins with specificity to glucose, D-fructose, D-mannose, sucrose, and methyl-β-D-glycopyranoside, while the chitin matrix is specific to N-acetyl-D-glycosamine and oligosaccharides [51,52].…”

Section: Gymnopilus Spectabilis (Gsl)mentioning

confidence: 99%

“…Thus, the formation of spacer arms minimizes or solves the related problem by expanding the possibilities of interaction, since it increases accessibility to all the binding centers available in a protein. In contrast, spacer arms can promote the non-specific interactions of various proteins on the matrix [51,76].…”

Section: Monolithic Polymeric Cryogelsmentioning

confidence: 99%

“…In this method, a monolith containing reactive epoxy groups (epoxy-activated monolith) is initially converted to the amine form (amine-activated monolith) by a reaction of epoxy groups with reagents containing amine grouping, such as ethylenediamine. The amine-activated monolith then reacts with glutaraldehyde, forming an aldehyde-activated monolith capable of reacting with an amine grouping of molecules [13,51]. Lysozyme, also known as N-acetylmuramide glycanohydrolase, is an enzyme that aids digestion.…”

Section: Monolithic Polymeric Cryogelsmentioning

confidence: 99%

See 1 more Smart Citation

Lectin Purification through Affinity Chromatography Exploiting Macroporous Monolithic Adsorbents

et al. 2023

View full text Add to dashboard Cite

Growing medical, engineering, biochemical, and biological interest has led to a steady pace of research and development into polymeric monolithic structures with densely interconnected pores for purifying bio compounds. Cryogels, which are generated by freezing a reactive polymerization mixture, are highlighted due to their versatility and low relative cost as macroporous, polymeric, monolithic adsorbents. The conversion of cryogels into affinity adsorbents is one possible alternative to their optimal application. Some of the most often utilized supports for immobilizing particular ligands are monolithic columns manufactured with epoxy radicals on their surfaces. The purification of biomolecules with a high degree of specificity, such as lectins and glycoproteins with an affinity for glycosylated groups, has garnered interest in the use of fixed non-traditional beds functionalized with ligands of particular interest. The interaction is both robust enough to permit the adsorption of glycoproteins and reversible enough to permit the dissociation of molecules in response to changes in the solution’s pH. When compared to other protein A-based approaches, this one has been shown to be more advantageous than its counterparts in terms of specificity, ease of use, and cost-effectiveness. Information on polymeric, macroporous, monolithic adsorbents used in the affinity chromatographic purification of lectins has been published and explored.

show abstract

“…However, due to the low binding constant and large molecular size of hIgM, macroporous support materials like monoliths are preferably used during hIgM purification [17,21,22,25]. Cryogels have a great potential as an affinity separation tool for the isolation of large molecules such as deoxyribonucleic acid (DNA), antibodies, proteins and cells [32][33][34][35][36].…”

Section: Introductionmentioning

confidence: 99%

Poly(Hydroxyethyl Methacrylate) Immunoaffinity Cryogel Column for the Purification of Human Immunoglobulin M

Bakhshpour

Topçu

Bereli

et al. 2020

Gels

View full text Add to dashboard Cite

Human immunoglobulin M (hIgM) antibodies are considered as hopeful tools for diseases therapy. Therefore, chromatography approaches are used to purify hIgM with a single step. In this study, we prepared a poly(hydroxyethyl methacrylate) based immunoaffinity p(HEMA-I) cryogel column by using cyanamide to immobilize antihuman immunoglobulin on the p(HEMA) cryogel for purification of hIgM in aqueous solution and artificial human plasma. The characterization of the p(HEMA) cryogel column was performed by using a scanning electron microscope (SEM), micro-computerized tomography (µ-CT), Fourier transform infrared spectroscopy (FTIR), swelling degree and macro-porosity. Further, the optimizations of various parameters were performed such as, pH, ionic strength, temperature and concentration of hIgM in aqueous solutions. In addition, the Langmuir adsorption model was supported by experimental results. Maximum adsorbed amount of hIgM corresponded to 11.1 mg/g at pH 5.75 [morpholino ethanesulfonic acid (MES buffer)]. Our results indicated that the p(HEMA-I) cryogel column can be reused at least 10 times without significant loss in adsorption capacity. As a natural source, artificial human plasma was selected for hIgM adsorption and the purity of hIgM was evaluated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

show abstract

Immobilization of sugars in supermacroporous cryogels for the purification of lectins by affinity chromatography

Cited by 23 publications

References 52 publications

Characterization of bioactive compounds in the biomass of black locust, poplar and willow

Characterization of bioactive compounds in the biomass of black locust, poplar and willow

Lectin Purification through Affinity Chromatography Exploiting Macroporous Monolithic Adsorbents

Poly(Hydroxyethyl Methacrylate) Immunoaffinity Cryogel Column for the Purification of Human Immunoglobulin M

Contact Info

Product

Resources

About