Antibody-antigenic peptide interactions monitored by SPR and QCM-D. A model for SPR detection of IA-2 autoantibodies in human serum.
This work reports on a complementary use of surface plasmon resonance (SPR) and quartz crystal microbalance with dissipation monitoring (QCM-D) technologies to study interactions between a peptide antigen and polyclonal antibodies, in an experimental format suitable for diagnostic assays of autoimmune diseases. In the chosen model, a synthetic peptide from the juxtamembrane region of IA-2 (a type 1 diabetes associated antigen) was immobilized by an optimized chemical protocol applicable to both BIACORE and QCM-D sensors. A thorough study of the peptide immobilization was performed to optimize the signal-to-noise ratio using mixed self-assembled monolayers (SAM) on a gold surface. Introduction of polyethylene glycol (EG(6)) chains into mixed SAM layers and addition of an anionic surfactant to the human serum reduced non-specific binding without modifying the viscoelasticity properties of the layer. Under our conditions, the antibody SPR detection limit was determined to be 0.2 nM in diluted human serum. This value is in agreement with the reported rank distribution of IA-2 antibodies in diabetic patient sera. Label-free and real-time technologies such as SPR and/or QCM-D could be precious tools in future diagnostic assays.
Ayela
Cedric
C
CNRS UPR 800, Laboratoire d'Analyse et Architecture des Systèmes, 7 Avenue du Colonel Roche, Toulouse, France.
Roquet
Francoise
F
Valera
Lionel
L
Granier
Claude
C
Nicu
Liviu
L
Pugnière
Martine
M
D000595:Amino Acid Sequence; D000937:Antigen-Antibody Reactions; D001323:Autoantibodies / Q000097:blood*; D006801:Humans; D008969:Molecular Sequence Data; D011791:Quartz / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000379:methods*
D001323:Autoantibodies; C096322:ICA512 autoantibody; D011791:Quartz