Forum :: CM5 Lifetime


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This forum is intended for questions about kinetics, Surface Plasmon Resonance and the instruments related to these techniques.
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TOPIC: CM5 Lifetime

CM5 Lifetime 22 Dec 2015 16:40 #1

Hi,

Anybody knows what the lifetime for CM5, I mean how many experiments I can run using one CM5, specially working with small molecules, mine is dead after 120 injections (30 injection/cycle) so what do you think?

CM5 Lifetime 23 Dec 2015 16:16 #2

Hi,
You probably mean the lifetime of the ligand on the sensor chip.
It depends on the ligand. Antibodies are quite robust but some other proteins not. It also depends on the buffer and regeneration conditions used.
And the analyte: pure protein or compound (DMSO) or culture medium.

So it is hard to predict how long a ligand surface will be functional.

Regards
Arnoud

CM5 Lifetime 23 Dec 2015 17:08 #3

Thanks Arnoud,

Do you think using the CM5 chip with compounds (DMSO) will affect the lifetime of the ligand on the sensor chip?

The other question is: why we can't reload the ligand to chip surface again after it loses its activity; I mean doing another immobilization to the same surface?

Regards
Ahmed

CM5 Lifetime 24 Dec 2015 09:51 #4

Hi Ahmed,

1) If the DMSO will affect your ligand is dependent on the nature of the ligand. With low concentrations (<2 %) this will be probably not a real issue. But you have to test each ligand for stability. If you have a positive control you can inject this every 1-15 cycles to test the ligand integrity.

2) Theoretically this is possible since the first activation of the sensor chip will not activate all the carboxyl groups of the dextran matrix. But when you try a second activation you will also modify the existing ligand on the surface....
I never have seen anyone doing this.

If sensorchip cost are an issue you can try to recondition the surface.
You should be very careful when using this procedure and make sure the chip is still firmly attached to the holder since aggressive reagents are used.

Ref: Chatelier, R. C., Gengenbach, T. R., Griesser, H. J., et al.; A general method to recondition and reuse BIAcore sensor chips fouled with covalently immobilized protein/peptide. Analytical Biochemistry (229) 1: 112-118; 1995.

Regards

Arnoud
Last Edit: 25 Dec 2015 10:58 by Arnoud.

CM5 Lifetime 24 Dec 2015 17:46 #5

Thanks a lot Arnoud

Best Regards
Ahmed

CM5 Lifetime 20 May 2016 13:05 #6

Hello,
What about storing an immobilized chip?
We used to store them wet, but we have had some trouble
because the Au film has been detached from the chip surface and
attached to the instrument,

Does anyone recommend another way to store the chips, we use XanTEC?

Thank you,
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