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CD3: And Another Thing...

We at UCL Advanced Diagnostics were lucky enough to be involved in the external assessment of CD3, LN10 in its validation stage. This enabled us to begin using it routinely as soon as it was commercially available, indicating our feelings on it! During the development and validation of CD3, LN10, we followed a similar testing regime to Professor Anagnostopoulos at the Charité Institute, Berlin.

The antibody was first optimised on reactive tonsil (see Figure 1) and subsequently stained on a variety of cases including T and B cell acute lymphoblastic lymphomas (Figures 2 & 3), peripheral T cell lymphomas (Figure 4), cutaneous T cell lymphomas (Figure 5) and nasopharyngeal NK/T cell lymphomas (Figure 6).

Prior to trying LN10, we were using another commercially available CD3 antibody (monoclonal) and during validation LN10 was compared to a number of other CD3 antibodies from various comm- ercial sources. In agreement with Professor Anagnostopoulos we also found LN10 to produce superior localisation and sensitivity to other CD3 antibodies on the market.

Using the Bond-maxTM automated immunostainer, we have optimised LN10 using ER2 (pH 8.0) for 30 minutes, followed by incubation with the primary antibody (1/200) for 20 minutes. The detection kit used was the Bond Polymer Refine Detection (DS9800) – incubation with post primary for 15 minutes, polymer for 15 minutes, DAB for 10 minutes and DAB enhancer for 5 minutes.

We now use LN10 routinely and a reflection of its efficacy is that we are consistently achieving excellent scores on our NEQAS assessments.

Figure 1. LN10 staining on reactive tonsil
Figure 2. LN10 staining on a T-ALL
Figure 3. LN10 staining on a B-ALL
Figure 4. LN10 staining on a peripheral T-cell lymphoma
Figure 5. LN10 staining in a cutaneous T-cell lymphoma
Figure 6. LN10 staining in a NK/T-cell lymphoma