Conjugation reactions in the preparations of quantum dot-based immunoluminescent probes for analysis of proteins by capillary electrophoresis

• Published in: Analytical and bioanalytical chemistry 2011-04, Vol.400 (2), p.369-379
• Main Authors: Lišková, M., Voráčová, I., Klepárník, K., Hezinová, V., Přikryl, J., Foret, F.
• Format: Article
• Language: English
• Subjects: Analytical Chemistry; Antibodies; Antibodies - chemistry; Antigens; Biochemistry; Capillarity; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Crosslinked polymers; Crosslinking; DNA; Electrophoresis; Electrophoresis, Capillary; Food Science; Immunoassay - instrumentation; Immunoassay - methods; Investigations; Laboratory Medicine; Lectins; Luminescent Measurements - instrumentation; Luminescent Measurements - methods; Molecular Probes - chemistry; Monitoring/Environmental Analysis; Original Paper; Proteins; Proteins - analysis; Proteins - isolation & purification; Quantum Dots; Reactions (nuclear)
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-011-4700-5

A number of biologically important molecules, such as DNA, proteins, and antibodies, are routinely conjugated with fluorescent tags for high-sensitivity analyses. Here, the application of quantum dots in the place of bright and size-tunable luminophores is studied. Several selected bioconjugation reactions via zero-length cross-linkers, long-chain linkers, and oriented methods for linking of quantum dots with proteins were tested. Anti-ovalbumin, anti-proliferating cell nuclear antigen, anti-hemagglutinin, and anti-CD3 membrane protein as model antibodies and annexin V were used as high-specificity selectors. The reaction yield and efficiency of the prepared immunoluminescent probes were tested by capillary zone electrophoresis with laser-induced fluorescence detection. Figure Scheme of antibody-quantum dot conjugate