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Critical steps in preparation of affinity immunosorbents.

Authors: Slováková Marcela | Švehlová Lucie | Jankovičová Barbora | Bílková Zuzana

Year: 2007

Type of publication: článek ve sborníku

Name of source: Amino Acids

Publisher name: Springer

Page from-to: LI

Titles:

Language	Name	Abstract	Keywords
cze	Kritické kroky při přípravě afinitních nosičů.	Nová diagnostická metoda založená na biochemických markerech Alzheimerovy choroby, peptidy A-beta a protein Tau. Příprava nosičů s imobilizovanými protilátkami anti-A-beta a anti-Tau. POužití magnetických nosičů. References: 1.?Purification of anti-chymotrypsin antibodies for the preparation of a bioaffinity matrix with oriented chymotrypsin as immobilized ligand?. Z. Bílková et al. Journal of Chromatography B, 689 (1997) 273-279. 2.?ImmunoAffinity Reactor for Prion Protein Qualitative Analysis?, Z. Bílková et al., Proteomics 5 (2005) 639. 3.?Immobilized magnetic enzyme reactors: effective tool for isolation and study of highly heterogeneous glycoproteins?, L. Korecká et al., J. of Magnetism and Magnetic Materials 293 (2005) 349. 4.?Use of self assembled magnetic beads for protein on-chip digestion?, M. Slováková et al.:, Lab-on-chip 5(9) (2005), 935-42.	afinitní nosiče, bioafinitní chromatografie, biochemické markery, Alzhemerova choroba, magnetické nosiče
eng	Critical steps in preparation of affinity immunosorbents.	New diagnostic methods reliant on biochemical biomarkers of disease hold the potential to provide effective measures of biological activity and markers that account for clinical benefit of the therapy. Detection of biomarkers is often based upon the use of specific antibodies immobilized on some type of support. The significance level of bioaffinity sorbents is in their versatile and potential utilization. This principle of bioaffinity chromatography was used in many applications, i.e. isolation and purification of biomolecules1, detection and identification of biomarkers2, selective decontamination of mixtures and use of immobilized enzymes for modification of target biomolecules3. We focus on neurodegenerative Alzheimer disease and its possible markers, especially the immunospecific supports dedicated to capturing and pre-concentration of target A peptides and Tau proteins. The goal in the construction of an efficient immunosorbent is to immobilize the antibody to the solid-phase support without adversely affecting the antibody?s function to capture antigen. The choice of support plays an important role, generally low non-specific sorption and chemical and physical stability are preferred. Rapid and simple manipulation and possible application into microfluidic device are facilitated by using magnetic supports4. The aim is in particular high steric accessibility of the active sites, high operational and storage stability of binding activity, and low non-specific sorption of protein to minimize the contamination of the target antigen. Therefore, suitable immobilization method of the specific anti-A-beta and anti-Tau antibodies with use of newly developed magnetic and non-magnetic carriers with controlled non-specific sorption will be described. In addition, new sights in quantification of isolated markers will be implemented. References: 1.?Purification of anti-chymotrypsin antibodies for the preparation of a bioaffinity matrix with oriented chymotrypsin as	affinity immunosorbents; bioaffinity chromatography; biochemical markers; Alzheimer´s disease; magnetic support