Site-specific conjugation of metal carbonyl dendrimer to antibody and its use as detection reagent in immunoassay. - Archive ouverte HAL Access content directly
Journal Articles Analytical Biochemistry Year : 2010

Site-specific conjugation of metal carbonyl dendrimer to antibody and its use as detection reagent in immunoassay.

Nathalie Fischer-Durand
Michèle Salmain
Bogna Rudolf
  • Function : Author
Lili Dai
  • Function : Author
Lauriane Jugé
  • Function : Author
Anne Vessières
Gérard Jaouen

Abstract

We describe here the conjugation of polyclonal goat anti-rabbit antibody to generation 4 polyamidoamine (G4-PAMAM) dendrimers carrying (i) (η(5)-cyclopentadienyl) iron dicarbonyl succinimidato complexes as infrared (IR) probes, (ii) nitroaniline entities as nuclear magnetic resonance (NMR) probes, (iii) acetamide groups for surface neutralization, and (iv) hydrazide-terminated spacer arms for the reaction with aldehyde. To preserve a high binding affinity, the conjugation was performed on the carbohydrate moieties located on the Fc fragment. The resulting conjugates were characterized by Fourier transform-IR, ultraviolet (UV), and high-mass matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. On the basis of relative concentration ratios of IR probes and antibody, an average labeling of 30 IR probes per antibody was reached (i.e., more than twice the value obtained with our previous strategy that generated no spacer arm). Immunoassays revealed that the antibody-dendrimer conjugates retained 55.1% of immunoreactivity on average with respect to underivatized antibody. Finally, the conjugates were used to quantify their antigen by solid-phase carbonyl metallo immunoassay (CMIA). Results showed a significant enhancement of the IR signal, demonstrating the efficiency of the new conjugation strategy and the potential of the new antibody-dendrimer conjugates as universal immunoanalytical reagents.

Dates and versions

hal-00534225 , version 1 (09-11-2010)

Identifiers

Cite

Nathalie Fischer-Durand, Michèle Salmain, Bogna Rudolf, Lili Dai, Lauriane Jugé, et al.. Site-specific conjugation of metal carbonyl dendrimer to antibody and its use as detection reagent in immunoassay.. Analytical Biochemistry, 2010, 407 (2), pp.211-9. ⟨10.1016/j.ab.2010.08.027⟩. ⟨hal-00534225⟩
197 View
0 Download

Altmetric

Share

Gmail Facebook X LinkedIn More