lifesciences
Innopsys

Carboxybetaine Modified Interface for Electrochemicla Glycoprofiling of Antibodies Isolated from Human Serum

Abstract:

Impedimetric lectin biosensors capable of recognizing two different carbohydrates (galactose and sialic acid) in glycans attached to antibodies isolated from human serum were prepared. The first step entailed the modification of a gold surface by a self-assembled monolayer (SAM) deposited from a solution containing a carboxybetaine-terminated thiol applied to the subsequent covalent immobilization of lectins and to resist nonspecific protein adsorption. In the next step, Sambucus nigra agglutinin (SNA) or Ricinus communis agglutinin (RCA) was covalently attached to the SAM, and the whole process of building a bioreceptive layer was optimized and characterized using a diverse range of techniques including electrochemical impedance spectroscopy, cyclic voltammetry, quartz crystal microbalance, contact angle measurements, zeta-potential assays, X-ray photoelectron spectroscopy, and atomic force microscopy. In addition, the application of the SNA-based lectin biosensor in the glycoprofiling of antibodies isolated from the human sera of healthy individuals and of patients suffering from rheumatoid arthritis (RA) was successfully validated using an SNA-based lectin microarray. The results showed that the SNA lectin, in particular, is capable of discriminating between the antibodies isolated from healthy individuals and those from RA patients based on changes in the amount of sialic acid present in the antibodies. In addition, the results obtained by the application of RCA and SNA biosensors indicate that the abundance of galactose and sialic acid in antibodies isolated from healthy individuals is age-related.

Credits:

Tomas Bertok1, ○ Alena Šedivá1, o Jaroslav Filip1, Marketa Ilcikova2, Peter Kasak2, Dusan Velic3,4, Eduard Jane4, Martina Mravcová5, Jozef Rovenský6, Pavol Kunzo6, Peter Lobotka6, Vasilij Šmatko7, Alica Vikartovská1, and Jan Tkac1

  • 1. Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, 845 38, Slovak Republic
  • 2. Centre for Advanced Materials, Qatar University, Doha 2713, Qatar
  • 3. Department of Physical Chemistry, Faculty of Natural Sciences, Comenius University, Mlynska Dolina, Bratislava, 842 15, Slovak Republic
  • 4. International Laser Centre, Ilkovičova 3, Bratislava 841 04, Slovak Republic
  • 5. Laboratory of Human Endocrinology, Institute of Experimental Endocrinology, Slovak Academy of Sciences, Vlarska 3, Bratislava, 833 06, Slovak Republic
  • 6. National Institute of Rheumatic Diseases, Nábrežie I. Krasku 4, 921 12 Piešt’any, Slovak Republic
  • 7. Department of Sensors and Detectors, Institute of Electrical Engineering, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava, 841 04, Slovak Republic

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