Can anyone recommend a way to make up Elution Blinds aside from using previously analyzed specimens?

If you haven't done it in a while (staff will catch on), check cells.

My old supervisor would antigen type a patient sample for something AHG, like Fya. Once we had patient sample that was Fya positive, she'd remove all the plasma, and add a ton of Anti-Fya sera (usually stuff that was about to expire). Let it sit in the fridge for several days so the Anti-Fya antisera bound to the patient cells that were Fya pos, then she'd check that the DAT was coming up pos at IgG, and make us do an eluate on that sample. 

The caveat here is that it's not a very stable sample, and it only worked well sometimes. Also, what I describe above comes from my memories of 10 years ago, so I may have missed a step. I haven't had to attempt to make this myself in my current facility. I need a whiz like @Malcolm Needs to check that this could work in theory......  

1 hour ago, jshepherd said:

My old supervisor would antigen type a patient sample for something AHG, like Fya. Once we had patient sample that was Fya positive, she'd remove all the plasma, and add a ton of Anti-Fya sera (usually stuff that was about to expire). Let it sit in the fridge for several days so the Anti-Fya antisera bound to the patient cells that were Fya pos, then she'd check that the DAT was coming up pos at IgG, and make us do an eluate on that sample. 

The caveat here is that it's not a very stable sample, and it only worked well sometimes. Also, what I describe above comes from my memories of 10 years ago, so I may have missed a step. I haven't had to attempt to make this myself in my current facility. I need a whiz like @Malcolm Needs to check that this could work in theory......  

I never did it like this.  We always incubated at 37oC, and usually had the luxury of using a mega amount of plasma from an entire unit of blood from an immunised donor, rather than from a patient or an expiring grouping reagent.  This also meant that we had the luxury of being able to chose a donor with a weak form of the antibody.  That worked for us (including preparing samples for teaching and exams at college/university.

Sadly, I retired in October 2016, so I am no longer in a position to even try the way your old supervisor did this.

Thank you so much for your kind words.  I am not at all sure that I deserve to be called "a whiz", but, being excessively vain, I'll take it!!!!!!!!!!  

2 hours ago, Pamo said:

Can anyone recommend a way to make up Elution Blinds aside from using previously analyzed specimens?

First step is to determine what are you trying to prove. Do you want staff to merely have a productive/reactive eluate, or do you need them to identify the specificity, too?

As Malcolm says, if you're in a blood center/centre, with access to whole units of plasma containing IgG antibodies of various specificities, it can be relatively easy to sensitize red cells with the appropriate phenotype (from your RBC inventory). Using D+ cells with anti-D is the simplest approach (or use Check Cells). This would certainly satisfy the mandate for a productive/reactive results. But, if you want the added challenge of antibody ID, using the same cocktail of reactants each time will lessen said challenge (as Cliff commented).

But, back to basics.....make sure you have a specificity which is IgG in nature, reactive only by IAT. It should be relative strong WITHOUT LISS or PEG enhancement - use the standard/original saline-IAT to chose your antibody source. Always incubate your sensitization phase at 37C. You may need to vary the ratios of packed cells to serum to get optimal results (typically at least 2 volumes of antibody to one volume of packed cells). The shelf life of the sensitized cells may be increased by suspension in a red cell storage solution/preservative.

There's a recipe for this in the AABB publication, Blood Banker Favorites: A Collection of the Best Recipes for Blood Sample Preparation.

Because the "test system" used to test an eluate is the same as an IAT, we do Direct Observation on the preparation of the eluate itself......and do blinds on other IAT techniques to make sure they get the right answer using those methods.