I intend to make a Coombs control which is not commercially available in my country.
Use serum of anyone having anti D in his/her blood (immunized during pregnancy or after mismatched transfusion). Incubate serum of this person with washed O Rh D pos red cells at 37oC for 10 min ( in ratio of 2:1 serum to cell). Wash cells and suspend them in CPD solution as 5 % suspension. Perform DAT on these cells which should be 2-3+. The cells will be good for 01 month. If want to extend life use cell stab of DiaMed or suspend them in Alsevers solution made in house.
Many years ago I used a chromium chloride coupling technique to produce monospecific IgG-, IgA- and IgM-coated cells as Coombs controls. I used serum from IgM Macroglobulinaemia, IgG and IgA myeloma patients to coat the cells. As Carlos says - some required diluting before use and needed testing with the other Coombs reagents for "non-specific" reactivity [i.e. testing IgG-coated cells with anti-IgA and -IgM. I seem to remember them lasting about three weeks, but that was ~30 years ago and the suspension media have improved a great deal since then. If I can find the method [no guarantee after all this time] would you like me to post it?
Thanks Graham! I won't be needing the technique. We decided to go for Gel Liss-Coombs technicque which does not require Coombs control.
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