US 2013/0281672 A1Published application

Chromatography Equipment Characterization

AssigneeHoffmann-La Roche Inc.

InventorsAnton Belousov, Thomas Dams, Benjamin Gerwat

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Abstract

Herein is reported a method for determining whether a re-usable chromatography column packing, which is used at least for the second time in a purification step of a purification of a polypeptide, has reduced separation efficacy in said purification step of said purification of said polypeptide, comprising the following steps: a) identifying and determining the experimental data of an inert change of at least one physicochemical parameter of a mobile phase passing through said re-usable chromatography column packing, b) determining the parameters of a function of formula I by fitting the experimental data of the inert change of the physicochemical parameter of the at least second use, c) determining the difference between the experimental data of the inert change of the physicochemical parameter of the at least second use and the function of formula I with the parameters determined in step b), d) calculating the difference between the maximum value and the minimum value of the difference determined in step c) and normalizing said difference, e) determining reduced separation efficacy of said re-usable chromatography column packing when the absolute value of the difference calculated in step d) is more than 0.1.

Claims14

Dependent claims

A method for determining whether a re-usable chromatography column packing, which is used at least for the second time in a purification step of a purification of a polypeptide, has reduced separation efficacy in said purification step of said purification of said polypeptide, characterized in that said method comprises the following steps:

a) identifying and determining the experimental data of an inert change of at least one physicochemical parameter of a mobile phase passing through said re-usable chromatography column packing of the at least second use,
b) determining the parameters of a function of formula I by fitting the experimental data of the inert change of the physicochemical parameter of the at least second use,
c) determining the difference between the experimental data of the inert change of the physicochemical parameter of the at least second use and the function of formula I with the parameters determined in step b),
d) calculating the difference between the maximum value and the minimum value of the difference determined in step c) and normalizing said difference,
e) determining reduced separation efficacy of said re-usable chromatography column packing when the absolute value of the difference calculated in step d) is more than 0.1,
wherein the function of formula I is
yI
=
-
1
2

A
·
exp

(
-
x
t
0
)

(
exp

(
2

x
c

t
0
+
w
2
2

t
0
2
)

(
erf
(
(
x
-
x
c
w
-
w
t
0
)
2
)
+
1
)
-
exp

(
x
t
0
)
·
erf

(
x
-
x
c
2
·
w
)
)
+
y
0
formula


I
with absolute signal values determined by the solvent employed (A and y0), the mean value and the standard deviation of the underlying Gaussian distribution function (xc, w), the time constant of the underlying exponential decay function (t0), and
wherein the erf-function used is defined as
erf

(
x
)
=
2
π

∑
0
∞

(
-
1
)
n

x
2

n
+
1
(
2

n
+
1
)

n
!
.

126 more elements

A method for the chromatographic purification of a polypeptide, wherein at least one chromatography step using a re-usable chromatography column packing is contained, characterized in that said method comprises the following steps:

a) identifying and determining the experimental data of an inert change of at least one physicochemical parameter of a mobile phase passing through said re-usable chromatography column packing of the at least second use,
b) determining the parameters of a function of formula I by fitting the experimental data of the inert change of the physicochemical parameter of the at least second use,
c) determining the difference between the experimental data of the inert change of the physicochemical parameter of the at least second use and the function of formula I with the parameters determined in step b),
d) calculating the difference between the maximum value and the minimum value of the difference determined in step c) and normalizing said difference,
wherein the function of formula I is
yI
=
-
1
2

A
·
exp

(
-
x
t
0
)

(
exp

(
2

x
c

t
0
+
w
2
2

t
0
2
)

(
erf
(
(
x
-
x
c
w
-
w
t
)
2
)
+
1
)
-
exp

(
x
t
0
)
·
erf

(
x
-
x
c
2
·
w
)
)
+
y
0
formula


I
with absolute signal values determined by the solvent employed (A and y0), the mean value and the standard deviation of the underlying Gaussian distribution function (xc, w), the time constant of the underlying exponential decay function (t0), and wherein the erf-function used is defined
erf

(
x
)
=
2
π


∑
0
∞

(
-
1
)
n

x
2

n
+
1
(
2

n
+
1
)

n
!
,
and
further using the re-usable chromatography column packing when the absolute value of the difference calculated in step d) is 0.06 or less, or
performing an additional characterization and/or assessment of the purified polypeptide when the absolute value of the difference calculated in step d) is more than 0.06 but less than 0.2, or
changing the re-usable chromatography column packing when the absolute value of the difference calculated in step d) is 0.2 or more.

128 more elements

Dependencies are inferred from the claim text.

Facts

Application: 13/922,101Filed 19 Jun 2013
Priority: EP 101962884.421 Dec 2010
Published: 24 Oct 2013
Term ends: Not estimatedThe source records don’t support an estimate.

Classification10 codes

G16C 20/30
Chemoinformatics, i.e. ICT specially adapted for the handling of physicochemical or structural data of chemical particles, elements, compounds…
C07K 1/22
By chromatography: affinity chromatography or related techniques based upon selective absorption processes
G01N 30/86
Column chromatography: signal analysis
G01N 30/8665
Signal analysis: for calibrating the measuring apparatus