Slajd 1

1
Interaction between glycoproteins and lectins
studied using AFM
Kateryna Lebed, Joanna Grybos, Grazyna
PykaFosciak, Malgorzata Lekka, Jan Styczen The
Henryk Niewodniczanski Institute of Nuclear
Physics, Polish Academy of Sciences,
Radzikowskiego 152, 31-342 Kraków, Poland
1. PROTEIN PATTERNING
Adhesion force measurements
CASE a (Con A immobilized using polymeric
stamp)
Microcontact printing method as a method for firm
protein anchoring onto a surface without
affecting their activity.
The unbinding force for single molecular pair
Fa 105 2 pN
PDMS - polydimethylsilane stamps
CASE b (Con A immobilized without patterning)
2. MOTIVATION The formation of protein arrays
onto solid surfaces using microcontact printing
technique as it has many potential applications
including the development of advanced biosensors.
The unbinding force for single molecular pair
Fb 108 2 pN
3. ATOMIC FORCE MICROSCOPY
The calculated binding forces between
concanavalin A and carboxypeptidase Y was about
100 pN in both cases.
Adhesion maps
Patterned substrate with different chemical
domains can be identified using force
spectroscopy
topography
topography
4. MATERIALS
Carboxypeptidase Y (CaY)Concanavalin A (ConA)
adhesion map
adhesion map
The studied proteincarbohydrate interaction was
represented by concanavalin A (Con A) and
carboxypeptidase Y (CaY) pair. Measurements were
performed in liquid i.e. TBS buffer containing
1mM concentrations of Ca and Mn, at room
temperature.
Force distribution
5. RESULTS
ConA CaY force
AFM images PDMS stamps
ConA glutaraldehyde force
depth of holes 1.5 µm width of stripe 5 µm
depth of holes 1.5 µm diameter of circle 5 µm
6. CONCLUSIONS
Con A micropatterns
? Protein immobilization using microcontact
printing method was performed with good
reproducibility ? The calculated unbinding
forces between concanavalin A and
carboxypeptidase Y was about 100 pN for both
immobilization ways indicating that microcontact
printing technique did not change biological
activity of proteins ? Patterned substrate with
different chemical domains can be identified
using force spectroscopy
height of protein layer 10 nm width of
stripes 5.1 µm diameter of circle 5.1 µm