| 摘要(英) |
There are two drawbacks of Immunosensor Chip required to be
improved, one is the shelf life of microchip and the other is sensitivity
enhancement. For the peptide ligand design, we searched for the
hydrophobic patch around the bottom Fc region of antibody and analyzed
the charge distribution among the hydrophobic patch. On the other hand,
we designed another peptide ligands based on the electrostatic interaction
and hydrophobicity which bind strongly to the bottom Fc region of
antibody, as well. In this study, we focused on the Rabbit IgG as the
antibody and the prostate specific antigen (PSA) as the antigen. To find a
peptide ligand with high affinity to Rabbit IgG, we designed all the
possible peptide sequences according to the previous methods and
assisted with molecular dynamics simulation (MD). After immobilizing
peptide ligands onto gold chip surface, we measured the binding
capacities of Rabbit IgG, PSA, and the secondary antibody (2nd antibody)
on gold chip by surface plasmon simulation (SPR). Also, we could obtain
the recognition efficiency of Rabbit IgG to antigen and the orientation
factor by SPR measurement. Besides, on specificity test, we performed
the designed peptide ligands on Mouse IgG2a.The result revealed that a
peptide ligand exhibits high affinity and specificity to Rabbit IgG and has
well effect on the recognition efficiency to PSA antigen. Therefore,
according to the strategy we proposing on peptide ligand design, we can
definitely immobilize the antibody orientedly onto the sensor chip and
further enhance the sensitivity of antibody to antigen detection. |
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