how to prepare check cells ( coomb's control cells)?

[mrkeramati]

I want to prepare check cells ( coomb's control cells). I need a protocol for it. woul you mind sending it to me.

[David Saikin]

It used to be easy before the anti-D was monoclonal. You may have to tough it out and try various dilutions until you find the one that only reacts at antiglobulin phase.

[mla]

Ortho's high protein D will work. We place about 1 ml of O= packed rbcs in a 12X75 tube, fill the tube to about an inch from the top with saline, add about 10 drops of high protein D, incubate at 37 degrees for 30 minutes, wash 3-4 times. We usually aliquot this and use a new aliquot each day for a week. Good Luck.

[Changezi]

make the wash O+ cell suspension add a drop of Anti-D , incubate for 30min and ready to use

(put the prepration date ... weekly better)

[Mabel Adams]

Yes, O pos cells would surely work better.

[mrkeramati]

Ortho's high protein D will work. We place about 1 ml of O= packed rbcs in a 12X75 tube, fill the tube to about an inch from the top with saline, add about 10 drops of high protein D, incubate at 37 degrees for 30 minutes, wash 3-4 times. We usually aliquot this and use a new aliquot each day for a week. Good Luck.

Dear mlam,

thanks for your reply. I have a few question.

1. We dont have monoclonal Anti-D containing only IgG antibody. Is it possible to prepare check cells with Anti- D containing IgG+IgM?

2. are you use check cells from packed RBC prepared from the donor blood bags or blood prepared from the patints? if you use the last one do you add preservative to this aliquot.

3. Why you use from high protein D reagent?

Thanks,

[mrkeramati]

make the wash O+ cell suspension add a drop of Anti-D , incubate for 30min and ready to use

(put the prepration date ... weekly better)

Dear ahmed,

I was wondering if you could send me your protocol with details for me.

Thanks

[mla]

oops sorry about the o=, meant to say o+.

[Changezi]

Dear ahmed,

I was wondering if you could send me your protocol with details for me.

Thanks

did you mean SOP you need ??

[mrkeramati]

did you mean SOP you need ??

yes. if it is possible

[mla]

Dear mlam,

thanks for your reply. I have a few question.

1. We dont have monoclonal Anti-D containing only IgG antibody. Is it possible to prepare check cells with Anti- D containing IgG+IgM?

2. are you use check cells from packed RBC prepared from the donor blood bags or blood prepared from the patints? if you use the last one do you add preservative to this aliquot.

3. Why you use from high protein D reagent?

Thanks,

Basically we use the high protein D because it was the only thing that we could get to work and not elute off the cells by the end of the week. All of the clonals, would work for a day and then we had to make more. When we make the check cells, we usually pull some donor segs off of two or three different units of blood rather than use patient cells. The cells seem to hold up for a week.