Adsorption/Elution for A & B Subgroups

[johna]

Does anyone have a good method for performing these adsorption/elutions using monoclonal antisera? We perform maybe 3 or 4 a year using an old method that utilizes an outdated polyclonal anti-A,B which is kept frozen. The procedure involves testing the eluate at a room temp incubation period and following that up with a 37C incubation and Coombs reading. The results have been mixed and eventually we'll run out of the polyclonal reagent. Thanks in advance.

[Yanxia]

In China we will test the eluation at 4C , room temp and 37C Coombs. Because some anti-A and B is prefer cold temp, I think we should do it. I have found some weak A and B antigens react stronger at 4C with monoclonal antibody than at room temp.

[johna]

In China we will test the eluation at 4C , room temp and 37C Coombs. Because some anti-A and B is prefer cold temp, I think we should do it. I have found some weak A and B antigens react stronger at 4C with monoclonal antibody than at room temp.

Thanks Shily! But if you are using monoclonal anti-A,B for the adsorption the 37C/Coombs part would not be applicable since the Coombs sera would not react with murine antibody. This is in contrast to the human source anti-AB that many of us older members of the profession fondly remember.

[Yanxia]

Thanks Johna for your pointing out ! Please give me a chance to explain it.

We perform these adsorption/elutions using human antibodies not the monoclonal antibodies. As we know part of human anti-A and anti-B prefer 4C.

I said those things above to prove monoclonal antibodies prefer 4C.

Can I get the conclusion that part of polyclonal antibodies prefer 4C? So I think we should test it at this temp, too.

Sorry for my English!

[johna]

Thanks Johna for your pointing out ! Please give me a chance to explain it.

We perform these adsorption/elutions using human antibodies not the monoclonal antibodies. As we know part of human anti-A and anti-B prefer 4C.

I said those things above to prove monoclonal antibodies prefer 4C.

Can I get the conclusion that part of polyclonal antibodies prefer 4C? So I think we should test it at this temp, too.

Sorry for my English!

Your English is fine (But unfortunately my Chinese is not!). If you are using human source reagents I would continue to do what you are doing. The problem today is that as far as I know these reagents are no longer available commercially in the U.S..

[Yanxia]

Johna, the human plasma or serum which we used is self-made( sorry, I don't know if I can call it like this). When we found someone's antibody(healthy donor) titer is very high( often higher than 1024), we will conserve it as reagent. I don't know whether this kind of doing is legal in U.S..

[johna]

Johna, the human plasma or serum which we used is self-made( sorry, I don't know if I can call it like this). When we found someone's antibody(healthy donor) titer is very high( often higher than 1024), we will conserve it as reagent. I don't know whether this kind of doing is legal in U.S..

Sily,

I wouldn't say that it's illegal in the U.S. to do an adsorption/elution with donor or patient anti-A,B but before monoclonal antisera came on the scene commercial antisera was used for this testing. The reason being was that reagent antisera was known to have a very high titer. Thanks for your comments and I wonder if anyone else who frequents this board might be using a patient/donor anti-A,B for their adsorptions.