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Working
Instruction
ABIO-WIS-2011
Blood
Pasma
and
Serum
Collection,
Processing
and
Storage
AdeptBio
UG
(haftungsbeschrnkt)
Karl-Wiechert-Allee
76
30625
Hannover,
Germany
Table of contents Page 1 Aim of the working instruction ........................................................................................ 2 2 Responsibilities ................................................................................................................ 2 3 Materials and Methods ................................................................................................... 2 4 Disclaimer ........................................................................................................................ 3
(Signature)
(Signature)
(Signature)
Valid
since
28-July-2011
Version
1.0
Page
2
of
3
Working
Instruction
ABIO-WIS-2011
Blood
Pasma
and
Serum
Collection,
Processing
and
Storage
AdeptBio
UG
(haftungsbeschrnkt)
Karl-Wiechert-Allee
76
30625
Hannover,
Germany
1 Aim
of
the
working
instruction
The
collection
of
high
quality
samples
is
a
prerequisite
for
research
and
development
in
the
life
sciences
industries
as
it
is
in
academia
and
should
always
be
carried
out
under
standardized
conditions.
This
instruction
describes
all
steps
of
collection,
processing,
storage
and
transport
of
blood
plasma
and
serum
samples.
Blood
is
collected
into
serum-gel
tubes
with
coagulation
inducer
and
K2ETDA-containing
tubes
by
venipuncture
from
a
superficial
vein
of
the
cubital
region
to
obtain
serum
as
well
as
plateletpoor
plasma.
Plasma
samples
shall
be
free
of
haemolysis,
coagulation-activation
and
cellular
contamination
or
platelet
activation.
Labelling
of
sample
containers
has
to
be
performed
in
a
way
that
allows
an
unambiguous
identification
of
the
sample.
2 Responsibilities
AdeptBio
provides
this
working
instruction
to
its
partnering
collection
sites
and
upon
request
to
all
parties
who
are
interested
to
collect
blood
serum
and
plasma
samples
for
research
and
development
under
standardized
conditions.
The
collection
of
blood
samples
must
be
performed
by
authorized
medical
(laboratory)
staff
only.
3 Materials
and
Methods
1. Venipuncture
from
a
cubital
vein
is
performed
using
a
20
or
21
gauge
needle
(e.g.
21
g
x
0.75
Butterfly
Safety-Lok
Needle
BD
#
367281
or
similar
CE-marked
product).
If
a
tourniquet
is
applied,
it
should
not
remain
in
place
for
longer
than
1
minute
(risk
of
falsifying
results
due
to
hemoconcentration).
As
soon
as
the
blood
flows
into
the
container,
the
tourniquet
has
to
be
released
at
least
partially.
When
more
time
is
required,
the
tourniquet
has
to
be
released
so
that
circulation
resumes
and
normal
skin
colour
returns
to
the
extremity.
Aspirate
blood
into
serum
containers
first
(e.g.
9
mL
Serum
Clot
Activator,
Gel
Separator
Vacuette
Tube,
CE
Greiner
#
455010
or
similar
CE-marked
product).
Depending
on
size
of
tube
available
or
volume
requested,
multiple
tubes
may
be
used.
Afterwards,
blood
is
drawn
into
standard
K2EDTA
containing
tubes
(e.
g.
9.0
ml
K2
EDTA
Vacuette
Tube,
CE
Greiner
#
455036
or
similar
CE-marked
product).
Depending
on
size
of
tube
available
or
volume
requested,
multiple
tubes
may
be
used.
Free
flow
with
mild
aspiration
has
to
be
assured
to
avoid
haemolysis.
Content
of
tubes
is
gently
mixed
by
slowly
inverting
all
tubes
10
times.
2. After
venipuncture
and
blood
draw:
Plasma
is
obtained
from
K2EDTA
tubes
by
centrifugation
for
10
min
at
2,000
x
g
at
room
temperature.
Preferentially
use
a
centrifuge
with
swing-out
rotor
applying
no- brake
protocols.
This
centrifugation
shall
be
started
immediately
after
blood
Valid
since
28-July-2011
Version
1.0
Page
3
of
3
Working
Instruction
ABIO-WIS-2011
Blood
Pasma
and
Serum
Collection,
Processing
and
Storage
AdeptBio
UG
(haftungsbeschrnkt)
Karl-Wiechert-Allee
76
30625
Hannover,
Germany
collection. The resulting plasma sample has now been separated from red and white blood cells in an efficient and gentle way. Nevertheless, a significant number of platelets (~25%) is still present. If platelet-poor plasma is requested an additional step of preparation can be applied as follows. Otherwise proceed and transfer the supernatant in aliquots of 1.0 mL into 2 mL cryovials. Be aware not to touch the buffy coat with the pipet tip during transfer. Optional second centrifugation step for platelet-poor plasma: The plasma sample from the previous centrifugation is transferred into a second vial for another centrifugation for 15 min at 2,500 x g at room temperature. After this centrifugation, the supernatant is transferred in aliquots of 1.0 mL into 2 mL cryovials. Serum is obtained by allowing the Serum tubes to stand upright at room temperature (23-25C) for 30 minutes to allow clotting mechanisms to fully function. Centrifuge at room temperature at 3,000 x g. Preferentially use a centrifuge with swing-out rotor applying no-brake protocols. After centrifugation, the supernatant is transferred in aliquots of 1.0 mL into 2 mL cryovials. 3. Samples are transferred to a 80 C ultrafreezer. Maximum processing time should be under 30 min. for plasma and under 60 min for serum. Storage is at 80C. 4. Transport of samples is performed on dry ice. According to the local situation in hospitals and private practices, adaptations of this working instruction may be necessary. 4 Disclaimer This working instruction is designed to provide optimal blood specimen collection, preparation and storage for general research and development purposes. It was initially developed for analysis of proteins, Peptides and other small molecules present in the fluid. We cannot take responsibility for the usefulness of this working instruction for other purposes. This working instruction is subject to change without prior notice.