Hello all,
We have begun to see a drop in the performance of some of our in house produced
antibodies. In one case the Ab was produced and affinity purified in 1997 and
stored at -80C at 1mg/ml in PBS and now is no longer efficacious. This is
concerning since we have a number of other Abs that are stored similarly. I’ve
been looking, unsuccessfully, for a publication that has analyzed the shelf
life of antibodies. If you know of a publication or have your own information
about storage life I’d be very happy if you would share that info.
Any feedback greatly appreciated.
Dear Brian,
we have observed the same behavior of antibodies who were stored for
only 4 years at -80°C. We have submitted data to the manufacturer but no
answer so far.
Hope you will receive some information since I was also looking for it
but was unable to find anything.
All the best
Goran
On 24-Feb-17 1:30 PM, Brian Hampton wrote:
> Hello all,
>
> We have begun to see a drop in the performance of some of our in house
produced antibodies. In one case the Ab was produced and affinity purified in
1997 and stored at -80C at 1mg/ml in PBS and now is no longer efficacious.
This is concerning since we have a number of other Abs that are stored
similarly. I’ve been looking, unsuccessfully, for a publication that has
analyzed the shelf life of antibodies. If you know of a publication or have
your own information about storage life I’d be very happy if you would share
that info.
Rest of post
>
> Any feedback greatly appreciated.
>
> Brian
> ――
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Hi Brian,
There is quite a bit of knowledge floating around on this subject.
Unfortunately I think you will find it is really Ab dependent so there are no
hard and fast rules. In addition one needs to rule out many variables (provide
more detail on storage) before arriving at a root cause. Sev. examples; the
rate of freezing and the head space and surface vol. matter. Can you guarantee
it hasn't gone through any freeze-thaw cycles, is the pI close enough to the pH
that upon freezing the change in pH has caused precipitation, any analytical
(SEC, gel, etc) tests performed to ensure an intact and competent molecule at
the correct concentration still exists in the vial. On and on. As an example
of generic shelf life most commercial sites list 24 months as the shelf life.
Also the functional assay used matters. In my experience, WB is not sensitive
to slight conformational changes as other assays.
Here are sev. good lit. refs. I have a ton more if interested.
https://www.labome.com/method/Antibody-Shelf-Life-How-to-Store-Antibodies.html
Biotechnol Prog. 2010 May-Jun;26(3):727-33. doi: 10.1002/btpr.377.
Impact of freezing on pH of buffered solutions and consequences for monoclonal
antibody aggregation.
Kolhe P1, Amend E, Singh SK.
J Histochem Cytochem. 1983 May;31(5):691-6.
Monoclonal antibody storage conditions, and concentration effects on
immunohistochemical specificity.
Ciocca DR, Adams DJ, Bjercke RJ, Sledge GW Jr, Edwards DP, Chamness GC, McGuire
WL.
Pharm Res. 2013 Apr;30(4):968-84. doi: 10.1007/s11095-012-0933-z. Epub 2012 Nov
27.
Effect of pH and excipients on structure, dynamics, and long-term stability of
a model IgG1 monoclonal antibody upon freeze-drying.
Park J1, Nagapudi K, Vergara C, Ramachander R, Laurence JS, Krishnan S.
Pharm Res. 2011 Apr;28(4):873-85. doi: 10.1007/s11095-010-0343-z. Epub 2011 Jan
7.
Frozen state storage instability of a monoclonal antibody: aggregation as a
consequence of trehalose crystallization and protein unfolding.
Singh SK1, Kolhe P, Mehta AP, Chico SC, Lary AL, Huang M.
Thanks Goran for your reply. In our case we actually produced them from
rabbits or as monoclonal Abs, so they are in house made. The one I refer to
below is apparently “dead” compared to a fresh commercial version of the same
Ab. We have other poly- and monoclonal Abs made in-house that I am worried
about.
At least in another case we can obtain fresh Ab from a commercial source that I
believe is made from our clone as a source of fresh to compare to what we have
stored for many years. Back then when these Abs were made, the term “rigor and
reproducibility” was not invented yet so no thought then was given to an assay
to use for testing and tracking efficacy of the Abs over time. Really the only
assay was whether a band of the right MW was or was not detected. Not ideal
for R&R.
The problem is that people generally have the idea that “antibodies are
forever” and so troubleshooting goes awry in many cases as a result.
Thanks,
Brian
> On Feb 24, 2017, at 8:46 AM, Goran Mitulović
<email obscured>> wrote:
>
> Dear Brian,
>
> we have observed the same behavior of antibodies who were stored for only 4
years at -80°C. We have submitted data to the manufacturer but no answer so
far.
>
> Hope you will receive some information since I was also looking for it but
was unable to find anything.
>
> All the best
>
> Goran
>
>
> On 24-Feb-17 1:30 PM, Brian Hampton wrote:
>> Hello all,
>>
>> We have begun to see a drop in the performance of some of our in house
produced antibodies. In one case the Ab was produced and affinity purified in
1997 and stored at -80C at 1mg/ml in PBS and now is no longer efficacious.
This is concerning since we have a number of other Abs that are stored
similarly. I’ve been looking, unsuccessfully, for a publication that has
analyzed the shelf life of antibodies. If you know of a publication or have
your own information about storage life I’d be very happy if you would share
that info.
Rest of post
>>
>> Any feedback greatly appreciated.
>>
>> Brian
>> ――
>> View topic http://list.abrf.org/r/topic/4ZUeBz5cs9tZ2bLXVn5Qwf
>> Leave group <email obscured>?Subject=Unsubscribe
>>
>
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>
>
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