Module 9 2021

22/03/2021

Surface Plasmon Resonance (SPR) Biacore platform - principle

reflected light

reflected light

regeneration

analyte

incident light

incident light

baseline

gold film

resonating surface plasmons

e-e- ee- e- - ee- - ee- - e-

ee- - ee-e- - ee- - ee-- e-

Y Y

flow

0

60 120 180 240 300 360 420 480 540 Time

SPR detects changes in mass in the aqueous layer close to a sensor chip surface by measuring changes in the refractive index. When molecules in the test solution bind to a target molecule the mass increases, when they dissociate the mass falls.

The Organisation for Professionals in Regulatory Affairs

23

Use of SPR - ADA detection & isotyping

GM-CSF hAb standard Slowly dissociating Ab

GM-CSF treated patient Rapidly dissociating Ab

RU

RU

time

time

isotyping

RU

Anti- hIgM

Anti-h IgG2

Patient serum

Anti- hIgG1

Anti- hIgG3

Anti- hIgG4

22000

1

2 3 4

1 2

3

4

21500

1 binding 2 amount bound 3 dissociation 4 regeneration

21000

20500

Amount bound

20000

Response

19500

Amount bound

19000

0

1200

1400

200

400

600

800

1000

s

Time

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24

12