Poisson Interval Distribution

and

Poisson Distributions
Random Walks discussion
by Joshua Wayment and
Ryan White
2/15/05

Poisson Interval Distribution

Single event process:
Probability of an event observed between t and t+dt
Divide time interval into large number of increments, n,
of length t/n.
0

t
n

2t 3t
n n

P(t;)= t//n

assume n>>1, gives greater

probability of only a single event

t t + dt

Probability an event does not occur in first increment

.= 1-t/n
Probability an event does not occur in n successive increments
= (1-t/n)n
Probability an event does occur in the next time increment dt
= dt

Limit n>>1, (1-t/n)n expansion

leads to power series e ! "t

Fig 6.10 Poisson Interval Distribution

P (t ; ! )dt = !e "!t dt

P(t;)

(Poisson interval distribution)

Expectation value of tn is

!
2

t n = # $t n e " $t dt =

n!
$n

Mean interval :

t =

1
!

Mean-square
interval:

t2 =

0
0

2t

3t

2
=2 t
!

Standard deviation:

(t

= t

! t

2 1/ 2

) = (2 t

! t

2 1/ 2

Poisson Distribution
Events in a given amount of time:
Probability of an event does not occur
0.35

t"

1 ! # $e !$t dt = e ! $t "

0.3

P(k;)

0.25

Probability an event occurs in interval t

0.2

0.15

0.1

t!

$e "$t 0 e " $ ( t !" t0 ) dt 0 = $t !e "$t !

0.05
0
0

10

Probability of two events

t!

Probability of k events:

$e

" $ t0

$ (t ! " t 0 )e

" $ ( t ! " t0 )

k !
P (k ; ) =
e
k!

dt 0 =

($t ! )2
2

e" $t !

frequency

Particle Capture Statistics

e " #!t (#!t ) k

p ( k , #!t ) =
k!
capture time (s)

-= -8.5

ln(p(0))

What it is

Ct = integration time
k = number of counts
= 1/ = average capture rate
ln( p (0, "!t )) = "!t

! = slope
= 0.98 s-1

What it should be

integration time (s)

Anti-biotin IGG binding to immobilized biotin molecules

Single molecule studies of antibody/ antigen binding.

Using TAMRA conjugated IgG we can investigate single
molecule kinetics for association and disassociation.
Meaning that instead of making an ensemble measurement
we are examining kinetics on a single molecule basis

Common IgG

For our purposes we use

monoclonal anti-biotin from
mice, conjugated with TAMRA

5-(and-6)-carboxytetramethylrhodamine
succinimidyl ester (TAMRA)

Binding kinetics (biphasic)

50

50

.25 g/ml

45
40
Data: Data1_B
Model: ExpDec2

30

Chi^2 = 2.83486
R^2
= 0.94187
y0
A1
t1
A2
t2

20

0
0
145.56279
0.04823
21.03456
0.33086

29.02043
0.0062
2.65471
0.0295

Binding Rate (s)

Number of Observations

40

kbind ~ 1.3 x 1010 M-1 s -1

35
30
25
20
15

kbind ~ 1.4 x 109 M-1 s -1

10
10
5

0.0

0.5

1.0

1.5

2.0

Time to bind (s)

Binding kinetics modeled using two

exponential system:

y=A1exp(-t1x)+ A2exp(-t2x)

0.5

1.5

2.5

3.5

[Ab] (nM)

What does it mean to have biphasic kinetics?

Anti-biotin has two possible routes of finding biotin
1) Diffusion through solution
2) Diffusion on surface

Unbinding kinetics
25

Number of Observations

40

Data: Data1_D
Model: ExpDec1

30

Chi^2 = 2.70126
R^2
= 0.9393
y0
A1
t1

20

0
0
173.5066
0.04749

16.45395
0.00232

10

Unbinding Rate (s)

.25 g/ml

20

15

10

0
0

0.0

0.2

0.4

0.6

0.8

1.0

0.5

y = A1exp(-t1x)

2.5

[Ab] (nM)

Time to Unbind (s)

Unbinding kinetics modeled using single

exponential

1.5

Unbinding kinetics
1) Simple one step process
2) Independent of concentration

3.5