Antibody ID

[Pamo]

How does one develop Antibody ID samples for training purposes?

[Cliff]

Hi @Pamo,

There are a few options.

• If you subscribe to a proficiency testing program, you can often save what they send you.  WARNING - do not use these samples until after you have received the results if you are a CLIA lab.
• You can spike your training samples with commercial antisera.  These don't always react as expected / hoped for.
• In my opinion, the best option is to save a recent prior patient sample that is no longer needed or expired.
• Commercial samples are also available, such as this from Hemo bioscience.

[Malcolm Needs ☆]

In the Reference Laboratory I worked in (in the UK, NHSBT-Tooting Centre in London), we would freeze any useful samples from both patients and donors (although the latter were quite rare), but we also belonged to a scheme named SCARF (Serum, Cells And Rare Fluids).  I must confess that, having retired in late 2016, I'm not certain that SCARF is still going, or, come to that, how much it costs to join.

One thing that I would caution against, and that is diluting a "strong" antibody to make a "weak" antibody (although this is far more important when trying to make a "weak antibody" to use as a control (for example, for an IAT), as a "strong" antibody has a completely different equilibrium constant (although this may not be too important if you just use them for teaching).

[Yanxia]

5 hours ago, Malcolm Needs said:

One thing that I would caution against, and that is diluting a "strong" antibody to make a "weak" antibody (although this is far more important when trying to make a "weak antibody" to use as a control (for example, for an IAT), as a "strong" antibody has a completely different equilibrium constant (although this may not be too important if you just use them for teaching).

May you kindly give us more details about it?  Is it about the ionic strength? Thank you.

[Malcolm Needs ☆]

13 hours ago, Yanxia said:

May you kindly give us more details about it?  Is it about the ionic strength? Thank you.

I confess that I am no expert on the chemistry of this, but, as I understand it from the late Prof. Patrick Mollison's book, it is purely to do with differences in the equilibrium constants of weak and strong antibodies.

[John C. Staley]

Not many have my option.  My wife has an Anti-D, an Anti-K and for a short time a detectible Anti-s.  I would just draw some of her blood every so often.  

[jnadeau]

I take plasma from group O patient samples (no longer needed) who have a negative antibody screen and add about 2 ul of antisera per 2 ml of plasma.  This usually will give 2-3+ reactions in gel (but as Cliff said it doesn't always act as expected).  I use expired 3% panel cells, negative for the antibody I spiked the plasma with, for them to use as a "patient".

[Malcolm Needs ☆]

22 hours ago, John C. Staley said:

Not many have my option.  My wife has an Anti-D, an Anti-K and for a short time a detectible Anti-s.  I would just draw some of her blood every so often.  

With a syringe and cannula I trust My Good Sir!!!!!!!!!!!!!!!!

[John C. Staley]

1 hour ago, Malcolm Needs said:

With a syringe and cannula I trust My Good Sir!!!!!!!!!!!!!!!!

Most assuredly!!  She's a nurse and would not allow such activity unless performed with the upmost professionalism!  

[Bet'naSBB]

HemoBioScience has some "Simulated Patient Plasmas" that work quite well - and "keep" past their expiration.  They even work when aliquoted and frozen.

We have, in the past, requested "Antibody Containing Plasmas" from our blood suppliers.  We use several different suppliers - some will and some won't save them for us.  Usually pay a nominal fee - but not nearly as much as manuf. antisera.  When we get them, we aliquot and freeze.

The least favorite way is  - when a patient has an antibody - or you ID a new antibody - we scavenge as many CBC's as possible, combine the plasmas and freeze.

 

We do have some dilute antisera - but as said before - often times does not work as expected.

[SbbPerson ☆]

On 3/7/2024 at 8:38 AM, Pamo said:

How does one develop Antibody ID samples for training purposes?

Our blood bank do not do titers. We send those out to a reference lab. We only do screens and AB ID's. Usually, we use our Anti-sera for training, for screens and ID's. We also have a few patients with very interesting samples(multiple Allos, Weak D, DARA, Anti-Ku, etc...), we save those and use them for training too. 

Edited March 15 by SbbPerson
spelling

[Sandi]

On 3/8/2024 at 10:54 AM, jnadeau said:

I take plasma from group O patient samples (no longer needed) who have a negative antibody screen and add about 2 ul of antisera per 2 ml of plasma.  This usually will give 2-3+ reactions in gel (but as Cliff said it doesn't always act as expected).  I use expired 3% panel cells, negative for the antibody I spiked the plasma with, for them to use as a "patient".

When I was an SBB student, (back in the day), our instructor showed us how to make positive antibody screen samples for a large group of students.  We would obtain an unused bag of plasma of each type and spike it with anti-D (could be used all semester and frozen for the next semester).  For the blind samples for each student, we would provide an Rh neg matching RBC to the plasma type. Then create a "panel" with Rh+ and Rh neg cells to match the profile of the antibody we wanted for that day. Of course, we had to check the reactivity at the beginning of each semester. Spiking with monoclonal didn't always work the same as when we had human Anti-D. Worked like a charm!!!

 

[Mabel Adams]

AABB published a book called Blood Bank Favorites: A Collection of the Best Recipes for Blood Sample Preparation.  It has instructions for about everything you might need to create.