ecd 0 Posted February 17 Share Posted February 17 I know we're a little late to this but I'm trying to write a procedure/validate using DTT to eliminate Dara interference using the premade 0.2M DTT from HemoBioscience and was wondering if anyone that's currently performing the testing would be willing to share their SOP. At first glance it seemed so simple: thaw, treat the cells, test them- done. I think we all know nothing is that simple though, so the more I work on it the farther down the rabbit hole I go (what temp does it thaw at, how important is a specific pH of 7.3 on this PBS really, how should I label this stupid stuff to keep people from thawing it too many times, etc). So if anyone is willing to let me look at how you're doing it I would really appreciate it. Thanks! Link to post Share on other sites
carolyn swickard 222 Posted February 18 Share Posted February 18 TS- Dithiothreitol -DTT- Treatment of RBCs.pdfThis is our procedure for the HemoBioScience product. it will be open for 30 days only. (I think) Don't worry about thawing it too many times - there is only 2-4 mls in each tube, so it doesn't last for that many pts. We have just thawed ours at room temp. We wrote the procedure using both the HemoBioScience procedureand the one in the AABB Tech Manual. Best of luck AMcCord and Henrique 1 1 Link to post Share on other sites
ecd 0 Posted February 19 Author Share Posted February 19 Thanks Carolyn! It's definitely helpful. Do you also use DTT to treat plasma to differentiate between IgG and IgM? I'm thinking since we're going to have the reagent we might as well add that to our options too but I'm not sure it's worth it. One step at a time I guess lol. Link to post Share on other sites
Cliff 443 Posted February 21 Share Posted February 21 Preparation and Testing of DTT Treated Plasma.docx AMcCord 1 Link to post Share on other sites
carolyn swickard 222 Posted February 22 Share Posted February 22 On 2/19/2021 at 7:28 AM, ecd said: Thanks Carolyn! It's definitely helpful. Do you also use DTT to treat plasma to differentiate between IgG and IgM? I'm thinking since we're going to have the reagent we might as well add that to our options too but I'm not sure it's worth it. One step at a time I guess lol. No - we do not have that procedure and I do not anticipate adding it. Link to post Share on other sites
NicolePCanada 27 Posted February 26 Share Posted February 26 Our DTT procedure is send it to a reference lab. I can't help out with this one. Sorry. AuntiS 1 Link to post Share on other sites
AuntiS 81 Posted March 4 Share Posted March 4 I'm also developing a procedure for DTT treated cells. Can I assume that the DTT treated cells can be used to help ID antibodies other than daratumumab without additional validation? I am intrigued by the use of DTT for differentiation between IgG and IgM antibodies but I don't think we will go forward with it here at our community hospital Link to post Share on other sites
Ensis01 124 Posted March 8 Share Posted March 8 0.2M DTT is used to treat red cells (deactivates/cuts Sulphur bonds: CD38 in DARA patients and several HTLA antibodies). 0.01M DTT is added to plasma in a process to differentiate between IgG and IgM antibodies. In my opinion using 0.2M DTT in a hospital setting for DARA patients makes sense WRT time and money. But the other uses of DTT (HTLA, IgG/IgM antibodies etc.) are too time consuming to justify, especially in a busy (or short staffed) hospital. AuntiS 1 Link to post Share on other sites
AuntiS 81 Posted March 8 Share Posted March 8 Thanks Ensis01! I haven't even gotten that far yet - so I didn't know that the concentration of DTT is different :) s Ensis01 1 Link to post Share on other sites
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