Benefits of immunomagnetic separation for epitope identification in clinically important protein antigens: a case study using ovalbumin, carbonic anhydrase I and tau protein

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00023752%3A_____%2F15%3A43914723" target="_blank" >RIV/00023752:_____/15:43914723 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216275:25310/15:39900558

Výsledek na webu

<a href="http://www.hrpub.org/journals/article_info.php?aid=2447" target="_blank" >http://www.hrpub.org/journals/article_info.php?aid=2447</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.13189/ujbe.2015.030101" target="_blank" >10.13189/ujbe.2015.030101</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Benefits of immunomagnetic separation for epitope identification in clinically important protein antigens: a case study using ovalbumin, carbonic anhydrase I and tau protein

Popis výsledku v původním jazyce

Immunomagnetic separation (IMS) with specific antibody as affinity ligand immobilized on a magnetic carrier has several advantages in comparison with standard column separation procedures. Epitope mapping enabling identification and characterization of protein structures reactive with the antibody represents one possible application of IMS. We used epitope extraction technique based on the proteolytic digestion of the target protein followed by capturing of a specific peptide fragments by the antibody immobilized on the solid phase. Magnetic particles coated with antibody molecules were first incubated with the prepared mixture of peptides. After specific binding of peptide fragments comprising the epitope sequences, the beads were washed to remove non-epitope peptides. Captured epitope-peptides were then eluted in small volume of 0.05% TFA. Elution fractions were finally analyzed without any modification by mass spectrometry. In this work the results and experience gained in epitope mapping of three clinically important proteins (ovalbumin, carbonic anhydrase I and tau protein) are discussed.

Název v anglickém jazyce

Benefits of immunomagnetic separation for epitope identification in clinically important protein antigens: a case study using ovalbumin, carbonic anhydrase I and tau protein

Popis výsledku anglicky

Immunomagnetic separation (IMS) with specific antibody as affinity ligand immobilized on a magnetic carrier has several advantages in comparison with standard column separation procedures. Epitope mapping enabling identification and characterization of protein structures reactive with the antibody represents one possible application of IMS. We used epitope extraction technique based on the proteolytic digestion of the target protein followed by capturing of a specific peptide fragments by the antibody immobilized on the solid phase. Magnetic particles coated with antibody molecules were first incubated with the prepared mixture of peptides. After specific binding of peptide fragments comprising the epitope sequences, the beads were washed to remove non-epitope peptides. Captured epitope-peptides were then eluted in small volume of 0.05% TFA. Elution fractions were finally analyzed without any modification by mass spectrometry. In this work the results and experience gained in epitope mapping of three clinically important proteins (ovalbumin, carbonic anhydrase I and tau protein) are discussed.

Druh

J_ost - Ostatní články v recenzovaných periodicích

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/ED2.1.00%2F03.0078" target="_blank" >ED2.1.00/03.0078: Národní ústav duševního zdraví</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2015

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Universal Journal of Biomedical Engineering

ISSN

2333-2662

e-ISSN

—

Svazek periodika

3

Číslo periodika v rámci svazku

1

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

8

Strana od-do

1-8

Kód UT WoS článku

—

EID výsledku v databázi Scopus

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