MP-SPR for Life Science

Why choose MP-SPR?

From lipids to living cells: MP-SPR enables to move from drug- target measurements, through drug-membrane interactions all the way to drug-cell interactions.

From Å to µm: Unique wide scanning angular range measurement ensures compatibility not only with thin layers (from Ångströms) but also thicker layers (up to a few micrometers). Re ected intensity From small to large molecules: Thanks to PureKinetics™, MP-SPR is a sensitive platform to determine drug-target interactions as well as nanoparticle-target interactions. Label-free interactions are measured in real-time revealing affinity and kinetics of the binding, whether the molecule is small or large. Indomethacin (358 Da) interaction with human serum albumin (HSA). Different concentrations of analyte (colour) are fitted (black curves) using TraceDrawer™ to obtain on- and off-rates as well as affinity. Functionalized gold nanoparticles (50 nm) interacting with a self-assembled polymer layer. Measured at 785 nm. Time (Seconds) (mdeg) CMD3D K D 1.39*10 -5 M k a 2.45*10 3 1/(M*s) k d 3.40*10 -2 1/s 20 -100 0 100 200 300 15 10 5 0 Angle (deg) 60 0,8 0,9 0,7 0,6 0,5 0,4 0,3 0,2 0,1 0 62 64 66 68 70 72 74 76 78 Re ected Intensity Calculated 1 Calculated 2 Calculated 3 Calculated 4 6,25 FC0 670nm 4,62 FC0 670nm 1,27 FC2 670nm 6,61 FC2 670nm

0,45

C

C

B

0,40

0,35

0,30

Deg

0,25

D

0,20

A

0,15

0,10

10

20

30

40

50

60

0

Time (min)

Lipid vesicles are bound to a hydrogel sensor surface and the interaction with a protein is studied. (A) Sensor cleaning injection, (B) Vesicle binding to surface, (C) Protein interaction, (D) Sensor regeneration.

Transcellular

Paracellular

0.5

0.5

D-mannitol

Propranolol

t = 0

0.0

0.0

t = 0

t = 1 hr

time

-0.5

-0.5

t = 1 hr

-1.0

-1.0

-1.5

-1.5

-2.0

-2.0

SP

SP

permeation

internalization

-2.5 -0.08 -0.06 -0.04 -0.02 0.00 0.02 0.04

-2.5 -0.08 -0.06 -0.04 -0.02 0.00 0.02 0.04

0.06

0.06

MP-SPR is the first label-free method that differentiates internalization from permeation. MP-SPR can also be used to study cell attachment on different coatings. Cells tested so far include HeLa, MDCKII, A549, LNCaP, ARPE19, PC-3, HepG2, MCF7, BK interacting with small molecules, nanoparticles including liposomes, silica, DNA polyplexes, viruses and microvesicles.

Not only function, but also structure: Thanks to multiple wavelengthsandLayerSolver™,MP-SPRhelpsyoutomeasurebiomembrane interaction kinetics as well as underlying structural changes. MP-SPR enables assessment of the lipid structure on the surface. Thickness and optical density of the layer shed light on the conformation.

0,95 0,15 0,2 0,25 0,3 0,35 0,4 0,45 0,5 0,55 0,6 0,65 0,7 0,75 0,8 0,85 0,9

0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 -0.1

EggPC+Label EggPC+POPS+Chol EggPC+POPS EggPC

Deg

39,4483 40

41

42

43

44

45

46

47

48

49

50

Angle

Single monolayer of graphene was measured as 3.5 Å thick at 670 nmwavelength. Thin layers form a single peak in a MP-SPR scan.

-20

0

20

40

Time (Minutes)

0.7

0.8

RLPO-PPZ RLPO-PVP-PPZ 4-spin RLPO-PVP-PPZ 2-spin

0.6

0.7

0.6

0.5

785 nm

Background Lipid bilayer

0.5

0.4

0.4

0.3

670 nm

0.3

0.2

Background Lipid bilayer

0.2

SPR angle (Deg) 0.1

Re ected intensity

0.1

0.0

0.0

0.1

58

60

62

64

66

68

70

72

74

76

-10

0

10

20

30

40

50

60

70

80

90

Angle (deg)

Time (Min)

Spreading of liposomes into supported lipid bilayers can be observed in real-time.

Perphenazine drug release from a micrometers-thick EUDRAGIT® polymer matrix. Faster release rates obtained by adding PVP polymer and varying thickness of the film.