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Preparation of antibody column

column antibody elution

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7 replies to this topic

#1 limpa1982

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Posted 10 August 2014 - 08:20 PM

Here is what I want to do. I want to coat columns with AAV2 and run serum through it. After that I want to collect the flow through and also want the elution through the column. How to coat the columns? How to do this procedure. Are there magnetic beads from any companies? Any protocol? Any name of this technique?  



#2 tkf

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Posted 11 August 2014 - 05:02 AM

I think you are looking to create an affinity column. I don't know what kind of AAV2 protein is, but you should find either BioRad Affi-gel 10 or Affi-gel 15  useful.

 

http://www.bio-rad.c...fi-gel-15-resin



#3 mdfenko

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Posted 11 August 2014 - 06:30 AM

you can use an antibody orientation kit from pierce (if AAV2 is an antibody as implied by the title of this thread).


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#4 limpa1982

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Posted 11 August 2014 - 11:03 AM

Thanks people. thats helpful. I actually AAV2 (adeno associated virus 2 serotype). I want to see if the the same antibodies of that bind to AAV2 also binds to AAV8. I am planning to coat the columns with AAV2 and then run the serum sample through it. How do I coat the columns with the virus I want?



#5 mdfenko

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Posted 11 August 2014 - 11:14 AM

you can bind it to cnbr-activated sepharose or ah- or ch-sepharose if you need an arm to hold the AAV@ away from the bead.

 

you can download a handbook of affinity chromatography from ge life sciences at this webpage: http://www.gelifesci...port/handbooks/

 

(as well as several other, useful handbooks).


Edited by mdfenko, 11 August 2014 - 11:15 AM.

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#6 limpa1982

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Posted 11 August 2014 - 11:40 AM

thanks



#7 limpa1982

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Posted 18 August 2014 - 07:43 PM

If I want to immobilize wav non-specifically on plate or column, how do I go about that?



#8 mdfenko

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Posted 20 August 2014 - 04:26 AM

If I want to immobilize wav non-specifically on plate or column, how do I go about that?

elisa plates will bind the protein non-specifically.

 

if you want to immobilize your protein, non-specifically, for a column then you can use cnbr activated agarose.

 

what do you mean by "non-specifically"? orientation?


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