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Bioaffinity magnetic isolation and preconcentration of specific antibodies from serum of patients with neurodegenerative disorders

Autoři: Slováková Marcela | Murasová Pavla | Vozandychová Věra | Jankovičová Barbora | Bílková Zuzana

Rok: 2018

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

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Jazyk	Název	Abstrakt	Klíčová slova
eng	Bioaffinity magnetic isolation and preconcentration of specific antibodies from serum of patients with neurodegenerative disorders	Development of efficient methods for fast, simple and selective isolation of specific antibodies of clinical or diagnostic significance remains an important challenge for researchers. An antigen immobilized to magnetic beads operated by magnetic separation can be gently and repeatedly used. Designing of the magnetic separation phase to be applied in a complex biological matrix as serum or cerebral fluid, requires outside the effective analytical system the low nonspecific sorption phase. More studies from the near past deal with the role of autoimmunity process in pathogenesis of Alzheimer disease (AD) [1-4]. In this work an occ urrence of the antibodies against the Amyloid beta 1-42 (anti- A) was observed, using the principle of magnetic bioaffinity chromatography. Preparation of the solid phase consisted of (1) an immobilization of Amyloid beta (A) 1-42 on various magnetic beads (1-3 µm) and (2) further steps for minimizing the contaminants nonspecific sorption. Five types of superparamagnetic beads varying in composition and size were characterized by scanning electron microscopy (SEM), dynamic light scattering (DLS) and zeta potential measurement. Activation of superparamagnetic beads was performed by carbodiimide chemistry followed by antigen immobilization. To block unoccupied beads functional groups the low molecular polyethylene glycol (PEG) agent was applied. Various techniques affecting the performance the non-specific sorption of the beads were evaluated using the anti-human IgG-peroxidase antibody (specific conjugate) and compared to the common blocking reactant, bovine serum albumin (BSA). Optimized magnetic bioaffinity solid phase was successfully used for preconcentration of the anti- A antibodies from serum of patients with neurodegenerative disorders.	Bioaffinity; magnetic; isolation; preconcentration; specific; antibodies; serum; patients; neurodegenerative; disorders

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