Open AccessUK NEQAS ICC & ISH Ki-67 Data Reveal Differences in Performance of Primary Antibody Clones
Author(s) -
Suzanne Parry,
Mitch Dowsett,
Andrew Dodson
Publication year - 2020
Publication title -
applied immunohistochemistry and molecular morphology
Language(s) - English
Resource type - Journals
eISSN - 1541-2016
pISSN - 1533-4058
DOI - 10.1097/pai.0000000000000899
Subject(s) - staining , antigen retrieval , confidence interval , clone (java method) , immunohistochemistry , antibody , medicine , pathology , biology , immunology , biochemistry , gene
We examined data from 374 laboratories staining for Ki-67 as part of external quality assessment over 8 runs between 2013 and 2017 (total data sets=2601). One of 5 primary antibodies was used for 94.8% of submissions, with MIB-1 (Agilent Dako) comprising 58.8% of the total. Examining assessment score as a continuous variable showed the 30-9 (Ventana) and K2 (Leica Biosystems) clones were associated with the highest mean scores (17.0; 95% confidence interval, 16.8-17.2 and 16.3; 95% confidence interval, 15.9-16.6, respectively). Stain quality was not significantly different between them. Both were associated with significantly better staining compared with MIB-1 (Agilent Dako), MM1 (Leica Biosystems), and SP6 from various suppliers (P<0.05). Similarly, categorical assessment of "Good" versus "Not good" staining quality showed that the 30-9 and K2 clones were both significantly associated with "Good" staining (both P<0.001). Other methodological parameters were examined for significant primary antibody-specific effects; none were seen for 30-9, K2, or SP6. The MM1 clone was more likely to be associated with good quality staining when it was used with Leica Biosystems sourced antigen retrieval, detection, and platform, all statistically significant at P<0.01. MIB-1 was more likely to be associated with good quality staining results when it was used with Agilent Dako antigen retrieval, detection, and staining platforms (P<0.0001), and less likely at the same significance level when used with Leica Biosystems reagents and equipment. The data presented here show the importance of not just primary antibody choice but also matching that choice to other methodological factors.