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

Autor: Merces AADD; Laboratório de Imunopatologia Keizo Asami, Departamento de Bioquímica, Universidade Federal de Pernambuco, Recife, Pernambuco, 50670-901, Brazil., Ferreira RDS; Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, São Paulo, 04044-020, Brazil., Silva KJS; Instituto Federal de Alagoas, Palmeiras dos Índios, Alagoas, 57608-180, Brazil; Centro de Ciências Exatas e da Natureza, Departamento de Física, Universidade Federal de Pernambuco, Recife, Pernambuco, 50670-901, Brazil., Salu BR; Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, São Paulo, 04044-020, Brazil., Maciel JDC; Centro de Ciências da Saúde, Universidade Federal de Roraima, Boa Vista, Roraima, 69310-000, Brazil., Aguiar JAO; Centro de Ciências Exatas e da Natureza, Departamento de Física, Universidade Federal de Pernambuco, Recife, Pernambuco, 50670-901, Brazil., Tashima AK; Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, São Paulo, 04044-020, Brazil., Oliva MLV; Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, São Paulo, 04044-020, Brazil., Carvalho Júnior LB; Laboratório de Imunopatologia Keizo Asami, Departamento de Bioquímica, Universidade Federal de Pernambuco, Recife, Pernambuco, 50670-901, Brazil. Electronic address: lbcj.br@gmail.com.
Jazyk: angličtina
Zdroj: Carbohydrate polymers [Carbohydr Polym] 2020 Nov 01; Vol. 247, pp. 116671. Date of Electronic Publication: 2020 Jun 22.
DOI: 10.1016/j.carbpol.2020.116671
Abstrakt: 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 Fe 2+ /Fe 3+ (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 Fe 3 O 4 in MAG-CH. This particle exhibited superparamagnetism and size between 100-300 μm. 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.
(Copyright © 2020 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: