Wojcik John B, Desai Keyur, Avraam Konstantinos, Vandebroek Arno, Dillon Lloye M, Giacomazzi Giorgia, Rypens Charlotte, Benci Joseph L
From Translational Sciences and Diagnostics (Wojcik, Dillon), Bristol Myers Squibb, Princeton, New Jersey.
Global Biometrics and Data Sciences (Desai), Bristol Myers Squibb, Princeton, New Jersey.
Arch Pathol Lab Med. 2023 Nov 1;147(11):1307-1314. doi: 10.5858/arpa.2022-0082-OA.
CONTEXT.—: An immunohistochemistry (IHC) assay developed to detect lymphocyte-activation gene 3 (LAG-3), a novel immune checkpoint inhibitor target, has demonstrated high analytic precision and interlaboratory reproducibility using a Leica staining platform, but it has not been investigated on other IHC staining platforms.
OBJECTIVE.—: To evaluate the performance of LAG-3 IHC assays using the 17B4 antibody clone across widely used IHC staining platforms: Agilent/Dako Autostainer Link 48 and VENTANA BenchMark ULTRA compared to Leica BOND-RX (BOND-RX).
DESIGN.—: Eighty formalin-fixed, paraffin-embedded melanoma tissue blocks were cut into consecutive sections and evaluated using staining platform-specific IHC assays with the 17B4 antibody clone. Duplicate testing was performed on the BOND-RX platform to assess intraplatform agreement. LAG-3 expression using a numeric score was evaluated by a pathologist and with a digital scoring algorithm. LAG-3 positivity was determined from manual scores using a 1% or greater cutoff.
RESULTS.—: LAG-3 IHC staining patterns and intensities were visually similar across all 3 staining platforms. Spearman and Pearson correlations were 0.75 or greater for interplatform and BOND-RX intraplatform concordance when LAG-3 expression was evaluated with a numeric score determined by a pathologist. Correlation increased with a numeric score determined with a digital scoring algorithm (Spearman and Pearson correlations ≥0.88 for all comparisons). Overall percentage agreement was 77.5% or greater for interplatform and BOND-RX intraplatform comparisons when LAG-3 positivity was determined using a 1% or greater cutoff.
CONCLUSIONS.—: Data presented here demonstrate that LAG-3 expression can be robustly and reproducibly assessed across 3 major commercial IHC staining platforms using the 17B4 antibody clone.
一种用于检测新型免疫检查点抑制剂靶点淋巴细胞激活基因3（LAG-3）的免疫组织化学（IHC）检测方法，在使用徕卡染色平台时已显示出高分析精度和实验室间的可重复性，但尚未在其他IHC染色平台上进行研究。
使用17B4抗体克隆评估LAG-3 IHC检测方法在广泛使用的IHC染色平台（安捷伦/达科自动染色仪Link 48和VENTANA BenchMark ULTRA）上的性能，并与徕卡BOND-RX（BOND-RX）进行比较。
将80个福尔马林固定、石蜡包埋的黑色素瘤组织块切成连续切片，并使用针对17B4抗体克隆的特定染色平台IHC检测方法进行评估。在BOND-RX平台上进行重复检测以评估平台内一致性。病理学家和数字评分算法使用数字评分评估LAG-3表达。使用1%或更高的临界值从手动评分中确定LAG-3阳性。
在所有3个染色平台上，LAG-3 IHC染色模式和强度在视觉上相似。当使用病理学家确定的数字评分评估LAG-3表达时，平台间和BOND-RX平台内一致性的Spearman和Pearson相关性均为0.75或更高。使用数字评分算法确定的数字评分时相关性增加（所有比较的Spearman和Pearson相关性≥0.88）。当使用1%或更高的临界值确定LAG-3阳性时，平台间和BOND-RX平台内比较的总体百分比一致性为77.5%或更高。
此处呈现的数据表明，使用17B4抗体克隆可在3个主要商业IHC染色平台上可靠且可重复地评估LAG-3表达。
