Identification of blood plasma proteins using heparin-coated magnetic chitosan particles

Author(s)

Dutra das Merces, Aurenice Arruda ^[1] ; Ferreira, Rodrigo da Silva ^[2] ; Santos Silva, Karciano Jose ^{[3, 4]} ; Salu, Bruno Ramos ^[2] ; Maciel, Jackeline da Costa ^[5] ; Oliveira Aguiar, Jose Albino ^[4] ; Tashima, Alexandre Keiji ^[2] ; Vilela Oliva, Maria Luiza ^[2] ; de Carvalho Junior, Luiz Bezerra ^[1]

Affiliation

^[1] Univ Fed Pernambuco, Dept Bioquim, Lab Imunopatol Keizo Asami, BR-50670901 Recife, PE - Brazil

^[2] Univ Fed Sao Paulo, Dept Bioquim, BR-04044020 Sao Paulo, SP - Brazil

^[3] Inst Fed Alagoas, BR-57608180 Palmeiras Dos Indios, Alagoas - Brazil

^[4] Univ Fed Pernambuco, Ctr Ciencias Exatas & Nat, Dept Fis, BR-50670901 Recife, PE - Brazil

^[5] Univ Fed Roraima, Ctr Ciencias Saude, BR-69310000 Boa Vista, Parana - Brazil

• Document type: Journal article
• Source: Carbohydrate Polymers; v. 247, NOV 1 2020.
• Web of Science Citations: 0

Abstract

Heparin was immobilized on magnetic chitosan particles to be used as a tool for human plasma protein identification. Chitosan was magnetized by co-precipitation with Fe2+/Fe3+ (MAG-CH). Heparin was functionalized with carbodiimide and N-hydroxysuccinimide and covalently linked to MAG-CH (MAG-CH-hep). X-ray diffraction confirmed the presence of chitosan and Fe3O4 in MAG-CH. This particle exhibited superparamagnetism and size between 100-300 mu m. Human plasma diluted with 10 mM phosphate buffer (pH 5.5) or 50 mM Tris-HCl buffer (pH 8.5) was incubated with MAG-CH-hep, and the proteins fixed were eluted with the same buffers containing increasing concentrations of NaCl. The proteins obtained were investigated by SDS-PAGE, LC/MS, and biological activity tests (PT, aPTT, and enzymatic chromogenic assay). Inhibitors of the serpin family, prothrombin, and human albumin were identified in this study. Therefore, MAG-CH-hep can be used to purify these proteins and presents the following advantages: low-cost synthesis, magnetic separation, ion-exchange purification, and reusability. (AU)

• FAPESP's process: 17/06630-7 - Fragments derived from the structure of protease inhibitors with selectivity for inhibition of mammalian and microorganism enzymes and its role as an anti-inflammatory, antimicrobial, antithrombotic and anti- tumor agent: mechanism of action
• Grantee: Maria Luiza Vilela Oliva
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 17/07972-9 - Development of lead agents for prophylaxis and treatment of cardiovascular diseases
• Grantee: Maria Luiza Vilela Oliva
• Support Opportunities: Regular Research Grants