Evaluation of autoantibody
In three videos Sue Johnson demonstrates the evaluation of a patient with a warm-reactive autoantibody including:
- The Direct Antiglobulin Test (22 min)
- Preparing and Testing an Eluate (25 min)
- Performing an Autoadsorption (30 min)
Again, details of proper technique are demonstrated. The videos were made at the Blood Center of Wisconsin, which kindly provided the laboratory facilities. They were shot and edited by Sharon Karp of Media Monster in Chicago, and produced and directed by Dr. Jim Perkins, director of the III…..by Indian immunohematology Initiative .
Thank you so much for your video on auto absorption I just want this we can replace ficin with bromiline if yes what will be the ratio with dtt
Thanks Sazia for your comment. Regarding your question, papain or ficin are usually used in ZZAP as mentioned in the AABB methods. However, I am not familiar with bromelain. I will inquire about it and get back to you.
Thank you for reply ma’am I performed all these tests but how can we validate our test . I performed all test just like your video but both were Panel reactive what should I do when my elution ID and autoadsorption is ID is panel reaction
Excuse me, I’m a man, not a woman.
I was asking ma’am in the video that’s why used female gender to talk you
This is Dr Don answer for this question ” In an attempt to answer your question regarding the use of bromelin in place of ficin for ZZAP reagent. I myself have only tested activated papain and ficin in combination with DTT in ZZAP formulations. Since both papain and ficin are proteolytic enzymes, as is bromelin, I would think that bromelin could be substitured for either papain or ficin. Best way to determine this is to compare ZZAP made with bromelin to ZZAP made with ficin and see if Kell system antigens are completely denatured. I would try and test Kell, cellano and Kpb if you have or can get antibodies to these Kell system antigens, but even just testing anti-K would be OK. Let me know if it works? Also, the pH with papain needed to be below 7.0 (liked pH 6.8 the best) for optimal activity of ZZAP (not published in original paper). So, if you can, check the pH when you make the ZZAP and try to have it at 6.8 if you can. My argument as to why W.A.R.M. doesn’t always work well for autologous adsorption is because the pH of W.A.R.M, is not optimal.”
Ok thanx I think I didn’t check ph at thr titof performing test
At the time of testing I mean
Thank you for reply ma’am I performed all these tests but how can we validate our test . I performed all test just like your video but both were Panel reactive what should I do when my elution ID and autoadsorption is ID is panel reaction
I used papain instead of ficin
As in AABB TM methods 20 ” ZZAP reagent by mixing 0.5 mL of 1% cysteine-activated papain with 2.5 mL of 0.2 M DTT and 2 mL of pH 7.3 PBS. Alternatively, use 1 mL of 1% ficin, 2.5 mL of 0.2 M DTT, and 1.5 mL of pH 7.3 PBS. ” Did you do that?
Ok thanx and regards
I did all process just like you only I use papain in zzap solution
Kindly answer my questions