Protein adsorption on histidyl-aminohexyl-Sepharose 4B: I. Study of the mechanistic aspects of adsorption for the separation of human serum albumin from its non-enzymatic glycated isoforms (advanced glycosylated end products) |
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| Authors: | Olivier Pitiot Laurent Folley Mookambeswaran A. Vijayalakshmi |
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| Affiliation: | 1. Institute of Mechanical Engineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais 1, 1049-001, Lisboa, Portugal;2. Department of Orthopedic Surgery, Henry Ford Health Systems, 2799 W Grand Blvd, Detroit, MI, 48202, United States;3. Division of Clinical Informatics, Beth Israel Deaconess Medical Center, 1330 Beacon St., Suite 400, Brookline, MA, 02446, United States;4. Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais 1, 1049-001, Lisboa, Portugal;5. Associate Laboratory I4HB—Institute for Health and Bioeconomy, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisboa, Portugal;6. Institute for Data, Systems and Society, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, United States;1. i3S-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen, 208, 4200-135 Porto, Portugal;2. INEB-Instituto de Engenharia Biomédica, Universidade do Porto, Rua do Campo Alegre 823, 4150-180-Porto, Portugal;3. INL-International Iberian Nanotechnology Laboratory, Avenida Mestre José Veiga s/n, 4715-330 Braga, Portugal;4. ICBAS-Instituto Ciências Biomédicas Abel Salazar da Universidade do Porto, Largo Prof. Abel Salazar, 2, 4099-003 Porto, Portugal;1. Institute of Quality Standards and Testing Technology for Agro-Products, Key Laboratory of Agro-Product Quality and Safety, Chinese Academy of Agricultural Sciences, Key Laboratory of Agri-Food Quality and Safety, Ministry of Agriculture and Rural Affairs, Beijing 100081, China;2. School of Materials Science and Engineering, Beijing Institute of Fashion Technology, Beijing 100029, China |
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| Abstract: | The characteristics of albumin adsorption on histidyl-aminohexyl-Sepharose 4B were investigated. In particular, the adsorption capacity of the gel was studied as a function of conductivity and pH of the running buffer. The adsorption was maximum at low salt concentration around neutral pH, involving electrostatic and hydrophobic interactions. Kinetic aspects were also investigated. Dissociation constant (KD) and maximum capacity (Qx) were, respectively, estimated to be 4.5×10−5 M (medium affinity) and 93.3 mg (high capacity) of human serum albumin per ml of adsorbent. According to these preliminary results, separation of HSA and its non-enzymatically glycated isoforms (conventionally named advanced glycated end products: AGEs) was achieved. Chromatographic potential of this separation tool is discussed. |
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| Keywords: | Proteins Histidyl-aminohexyl-Sepharose 4B Human serum albumin Advanced glycated end products |
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