Stability of Murine Monoclonal Anti‐A, Anti‐B and Anti‐A,B ABO Grouping Reagents and A Multi‐Centre Evaluation of Their Performance in Routine Use¹

Abstract

We have previously reported the production of 3 murine monoclonal reagents for ABO typing (designated ES-9, ES-4 and ES-15). This study presents results of tests of stability of these 3 reagents, together with a fourth murine monoclonal antibody (LM103/107). In addition, data are also presented from a multi-centre evaluation of the performance of the murine monoclonal reagents in routine ABO typing of both donors and patients using a wide variety of techniques, both manual and automated. The potency and stability of the 4 monoclonal antibody based reagents is compared with a broad selection of monoclonal and polyclonal ABO typing reagents. The reagents used for comparison were produced by European and United States manufacturers in both the public and private sector and are widely used in routine ABO typing. The Scottish monoclonal reagents have been used successfully to ABO type over 500,000 blood samples in 7 centres within the UK, with no discrepant results.

Mapping the fine specificity of ABO monoclonal reagents with A and B type‐specific function‐spacer‐lipid constructs in kodecytes and inkjet printed on paper

“…Since their introduction for routine serologic use in the early 1980s,1‐3 monoclonal antibodies (MoAbs) have improved the quality of ABO typing and created an unlimited resource of characterized reagents. The exact antigen structure used for primary immunization to generate MoAb clones is usually unknown, with heterogenous material including soluble antigens from ovarian cyst fluid and salivary glycoproteins or human cells, containing a range of ABH antigens being used. Selection of clones for serologic use is then usually based on direct agglutination specificity against red blood cells (RBCs), which also carry a heterogenous range of different types of ABO and related antigens.…”

A New Monoclonal Anti-A Antibody BIRMA-1

“…Thus the antibody titre of 256 after 4 months storage at 37°C indicates an extremely stable anti body. Scores obtained (table 6) are superior to those quot ed by McGowan et al [1] for a range of commercial re agents studied over an equivalent period. Surprisingly, we observed an increase in titre after 1 month storage at all temperatures.…”

“…Indeed, Messeter et al [8] discussed composing cocktails of anti-A antibodies which would recognise and distinguish all ABO variants, thus making anti-A,B reagents redundant. Workers are reluctant to give up the use of anti-A,B be cause of its confirmatory role in ABO typing, but never theless a single unblended monoclonal anti-A reagent that is both potent and able to detect Ax would be most useful [1]. To date we do not know of such a reagent, though some potent antibodies with reactivity for the Ax antigen have recently been reported in the proceedings of the Second International Workshop on Monoclonal Antibodies against Human Red Blood Cells and Related Antigens [12].…”

“…Ideally, such antibodies should have high avidity and po tency and be capable of detecting weak variants of the A antigen such as Ax. These characteristics are rarely shown by a single antibody and indeed, many of the com mercially available monoclonal anti-A reagents which fulfil these criteria are likely to be blends of more than one antibody [1].…”

Production of Stable Human-Mouse Hybridomas Secreting Monoclonal Antibodies against Rh D and c Antigens

“…Several years ago, murine monoclonal antibodies against A, B, AB, Lewis, Pi blood group antigens were produced [1][2][3][4]. They are now used worldwide in blood group laboratories.…”