Chip Design

  • Epitope (known) YPYDVPDYA
  • Chip (1) 800 HA and flag epitope and variant sequences
  • >3 redundancy
  • CV < 15%
  • Chip (2) HA, c-myc, VSVG, and flag epitope and other peptide sequences
  • >3 redundancy
  • CV < 30%

Assay

  • Antibody: anti-HA (100-1,000 ng/ml)
  • 200 µl TBS
  • Temp: 4oC
  • Reaction time: 1 hour

Detection

  • Cy5-IgG

Results

  • Figure 1 – Fluorescent image of Anti HA binding to epitope and its variants

Figure 1

  • Figure 2 – Plots of the signal intensities of the epitope sequence variants

Figure 2

YPYDVPDYARed: important residues in the 11-mer epitopeYPYDVPDYABlue: residues dispensable from both ends and give comparable or lower binding affinity.

Anti HA Binding: Association Curves Measured with Protein Concentration Titration on Epitope Peptide Chip Quantitative Measurement of Anti VSVG Epitope Peptide Binding