3104, PEG-2-128, PEG(backbone) - Epitomics

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Dec 14, 2013 (3 years and 6 months ago)

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For research use only. Not for use in diagnostic or therapeutic applications.

This product was manufactured under U.S. Patent No. 5,675,063. For a complete list of protocols and availab
le related products, please visit
us
at
www.epitomics.com.


Epitomics, Inc., 863 Mitten Road, Suite 103 Burlingame, California 94010
-
1303

Rev. A/0
4
-
05/JS











Clone ID: PEG
-
2
-
128
-
7

Lot #:

please refer to vial


Quantity:
100ug

Type:
Rabbit Monoclonal
IgM

Applications:

+
WB
+
ELISA
n/t
IHC

Concentration:
Please refer to vial



Background
:
P
olyeth
ylene glycol
ʢ
PEG
ʣ
is a family of long
chain polymers attached to a glycerine backbone. It is a
nonionic,
nontoxic,
biocompatible, strongly hydrophilic
polymer, which has a large exclu
sion volume in aqueous
solution (
1
).
The covalent attachment of PEG is now
commonly us
ed to modify a variety of proteins and drugs

(
2
,
3
).
The modification of a biopharmaceutical with
polyethylene glycol (PEG) increases its hydrodynamic radius
,

reduces immunogenicity
and proteolytic cleavage.
Other
benefits include decelerated renal excreti
on, improved
stability towards proteolysis and increased solubility of the
biopharm
aceutical in aqueous solutions (4).
As examples,
PEG
-
adenosine deaminase

(Adagen®) is used for the
treatment of severe combined immunodeficiency syndrome,
PEG

asparaginase


(Oncaspar®) is used for the treatment
acute lymphoblastic leukemia, PEG

interferon α2a
(Pegasys®) is used for the treatment Hepatitis C, Branched
PEG

anti
-
VEGF aptamer (Pegaptanib, Macugen™) is used
for the treatment Mac
ular degeneration(age
-
related) (5).


An
Anti
-
PEG antibody can be used to monitor
a drug’s
pharmacokinetics, including
distribution
,

metabolism and
excretion. In addition, it can be used for the quality control of
pegylated molecules in ELISA, WB and flow cytometer
y.


Background References
:

1. Guiotto, A. et al. (2004) Anchimeric assistance effect on
regioselective hydrolysis of branched PEGs: a mechanistic
investigation. Bioorg. Med.Chem. 12, 5031

5037

2. Wong, S.S. (1991) Reactive groups of proteins and their
modifying agents. In Chemistry
of protein conjugation and
cross
-
linking, p. 13, CRC Press

3. Caliceti, P. et al.(1993) Active site protection of proteolytic
enzymes by poly(ethylene glycol) surface modification. J.
Bioact. Comp. Polym. 8,41

50

4. Frank Leenders, celares GmbH, Berlin, G
ermany.(2006)
PEGylation technology and biopharmaceuticals.
Biopharmaceuticals. 6,39
-
40

5. Francesco M.Veronese, Gianfranco Pasut. (2005)
PEGylation, successful approach








Accuracy
: By detecting the backbone of the PEG molecule
itself, anti
-
PEG antib
ody is useful in
measuring the
pharmacokinetics o
f PEG
-
modified molecules. Data indicate
that this antibody detects PEGylated proteins as well as
PEG polymers.



Specificity
:
KLH
-
PEG20K was used as an immunogen. This
antibody recognizes the backbone o
f the PEG molecule.


Storage Conditions
:
Store at 4
°C
or aliquot and store at
-
20
°C. Buffer:
PBS, 0.01% sodium azide
. Stable for 12
months from date of receipt.


Recommended
Dilutions
:

WB:
1:1000

ELISA: 2
-
10 ug/ml


Please visit www.epitomics.com
for recommended protocols















Product QC’d by: __________________________________












Anti
-
P
EG
(backbone)
Rabbit Monoclonal Antibody

Product Data Sheet

Order: 877
-
7
7
2
-
2622

Support:
support@epitomics.com

Web: www.epitomics.com

Catalog #
3104
-
1


For research use only. Not for use in diagnostic or therapeutic applications.

This product was manufactured under U.S. Patent No. 5,675,063. For a complete list of protocols and availab
le related products, please visit
us
at
www.epitomics.com.


Epitomics, Inc., 863 Mitten Road, Suite 103 Burlingame, California 94010
-
1303

Rev. A/0
4
-
05/JS


Direct ELISA












































Sandwich ELISA

























Western Blot



















































Fig 3
.
1 ug (lane 1) and 0.5 ug (lane 2) of
MBP
-
PEG20K and 1 ug (lane 3) and 0.5 ug
(lane 4) of PEG20K were electrophoresed
on a 4
-
20% gel, transfe
rred to
n
itrocellulose
membrane, and probed with
anti
-
PEG
antibody
.

Fig 1a.

Various concentrations
of PEG polymers with amine end groups were
coat
ed in 96
-
wells. 2 ug/ml of anti
-
PEG
antibody was added. After washing, 1
ug/ml of anti
-
r
abbit IgM
-
HRP was incubated

Fig 2.

10 ug/ml of anti
-
PEG antibody (cat# 3104
-
1)
was coated into 96
-
wells. Serial dilution of PEG20K
conjugated MBP and PEGylated ADI were added.
After washing, 1 ug/ml of biotinylated anti
-
PEG
ant
ibody (cat. # 2137
-
1) was added. Streptavidin
-
HRP was then used to develop the color.


Fig 1b.
Various concentrations of PEG20K conjugated MBP and branched

PEG conjugated BSA were coat
ed in 96
-
wells. 2 ug/ml of anti
-
PEG
antibody was

added. After washing, 1 ug/ml of anti
-
ra
bbit IgM
-
HRP was incubated

50

75

100

150

250

1 2 3 4

0
0
.
5
1
1
.
5
2
2
.
5
1
1
0
1
0
0
1
0
0
0
n
g
/
m
l
OD450
M
B
P
-
P
E
G
2
0
K
A
D
I
-
P
E
G
0
0
.
5
1
1
.
5
2
2
.
5
3
3
.
5
4
0
.
1
1
1
0
1
0
0
n
g
/
m
l
OD450
M
B
P
-
P
E
G
2
0
K
B
S
A
-
b
r
a
n
c
h
e
d

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