PD-L1 expression in non-small cell lung cancer: evaluation of the diagnostic accuracy of a laboratory developed test using clone E1L3N in comparison with 22C3 and SP263 assays

Enrico Munari, Giuseppe Zamboni, Gianluigi Lunardi, Marcella Marconi, Matteo Brunelli, Guido Martignoni, George J Netto, Linda Quatrini, Paola Vacca, Lorenzo Moretta, Giuseppe Bogina

Research output: Contribution to journalArticle

Abstract

Different studies have evaluated the comparability of various immunohistochemical assays for PD-L1 expression evaluation, with contrasting results. Besides the important issues related to analytic performance and comparability of validated assays, not all platforms are available in all laboratories; moreover, standardized assays are very expensive and funding for PD-L1 testing is hard to obtain, especially in the research setting. One of the most widely used and inexpensive PD-L1 clone is E1L3N (Cell Signaling Technology, Danvers, MA), which is labeled for research use only. In this work, we wanted to further study and validate in a larger cohort the analytical performance of E1L3N clone on Ventana platform and its comparability with assays SP263 and 22C3 run onto their dedicated platforms. Serial sections of tissue microarrays built from 165 cases of resected lung cancer were stained for E1L3N onto Ventana platform following a previously reported protocol and for 22C3 and SP263 assays onto their respective platforms following manufacturer's instructions. Overall, we found very high concordance when comparing E1L3N with SP263 both at 1% and 50% cutoffs. Lower overall concordance was found between E1L3N and 22C3 at both cutoff; however, 100% sensitivity was found for E1L3N compared with both SP263 and 22C3 at 50% cutoff. Given the 100% sensitivity at 50% cutoff demonstrated by E1L3N in comparison with both SP263 and 22C3 and therefore the lack of false negative cases, we propose an algorithm for PD-L1 testing in NSCLC when considering pembrolizumab as first line therapy.

Original languageEnglish
JournalHuman Pathology
DOIs
Publication statusE-pub ahead of print - May 20 2019

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Non-Small Cell Lung Carcinoma
Clone Cells
Research
Lung Neoplasms
Technology
Therapeutics
pembrolizumab

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PD-L1 expression in non-small cell lung cancer : evaluation of the diagnostic accuracy of a laboratory developed test using clone E1L3N in comparison with 22C3 and SP263 assays. / Munari, Enrico; Zamboni, Giuseppe; Lunardi, Gianluigi; Marconi, Marcella; Brunelli, Matteo; Martignoni, Guido; Netto, George J; Quatrini, Linda; Vacca, Paola; Moretta, Lorenzo; Bogina, Giuseppe.

In: Human Pathology, 20.05.2019.

Research output: Contribution to journalArticle

Munari, Enrico ; Zamboni, Giuseppe ; Lunardi, Gianluigi ; Marconi, Marcella ; Brunelli, Matteo ; Martignoni, Guido ; Netto, George J ; Quatrini, Linda ; Vacca, Paola ; Moretta, Lorenzo ; Bogina, Giuseppe. / PD-L1 expression in non-small cell lung cancer : evaluation of the diagnostic accuracy of a laboratory developed test using clone E1L3N in comparison with 22C3 and SP263 assays. In: Human Pathology. 2019.
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abstract = "Different studies have evaluated the comparability of various immunohistochemical assays for PD-L1 expression evaluation, with contrasting results. Besides the important issues related to analytic performance and comparability of validated assays, not all platforms are available in all laboratories; moreover, standardized assays are very expensive and funding for PD-L1 testing is hard to obtain, especially in the research setting. One of the most widely used and inexpensive PD-L1 clone is E1L3N (Cell Signaling Technology, Danvers, MA), which is labeled for research use only. In this work, we wanted to further study and validate in a larger cohort the analytical performance of E1L3N clone on Ventana platform and its comparability with assays SP263 and 22C3 run onto their dedicated platforms. Serial sections of tissue microarrays built from 165 cases of resected lung cancer were stained for E1L3N onto Ventana platform following a previously reported protocol and for 22C3 and SP263 assays onto their respective platforms following manufacturer's instructions. Overall, we found very high concordance when comparing E1L3N with SP263 both at 1{\%} and 50{\%} cutoffs. Lower overall concordance was found between E1L3N and 22C3 at both cutoff; however, 100{\%} sensitivity was found for E1L3N compared with both SP263 and 22C3 at 50{\%} cutoff. Given the 100{\%} sensitivity at 50{\%} cutoff demonstrated by E1L3N in comparison with both SP263 and 22C3 and therefore the lack of false negative cases, we propose an algorithm for PD-L1 testing in NSCLC when considering pembrolizumab as first line therapy.",
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AU - Munari, Enrico

AU - Zamboni, Giuseppe

AU - Lunardi, Gianluigi

AU - Marconi, Marcella

AU - Brunelli, Matteo

AU - Martignoni, Guido

AU - Netto, George J

AU - Quatrini, Linda

AU - Vacca, Paola

AU - Moretta, Lorenzo

AU - Bogina, Giuseppe

N1 - Copyright © 2019. Published by Elsevier Inc.

PY - 2019/5/20

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AB - Different studies have evaluated the comparability of various immunohistochemical assays for PD-L1 expression evaluation, with contrasting results. Besides the important issues related to analytic performance and comparability of validated assays, not all platforms are available in all laboratories; moreover, standardized assays are very expensive and funding for PD-L1 testing is hard to obtain, especially in the research setting. One of the most widely used and inexpensive PD-L1 clone is E1L3N (Cell Signaling Technology, Danvers, MA), which is labeled for research use only. In this work, we wanted to further study and validate in a larger cohort the analytical performance of E1L3N clone on Ventana platform and its comparability with assays SP263 and 22C3 run onto their dedicated platforms. Serial sections of tissue microarrays built from 165 cases of resected lung cancer were stained for E1L3N onto Ventana platform following a previously reported protocol and for 22C3 and SP263 assays onto their respective platforms following manufacturer's instructions. Overall, we found very high concordance when comparing E1L3N with SP263 both at 1% and 50% cutoffs. Lower overall concordance was found between E1L3N and 22C3 at both cutoff; however, 100% sensitivity was found for E1L3N compared with both SP263 and 22C3 at 50% cutoff. Given the 100% sensitivity at 50% cutoff demonstrated by E1L3N in comparison with both SP263 and 22C3 and therefore the lack of false negative cases, we propose an algorithm for PD-L1 testing in NSCLC when considering pembrolizumab as first line therapy.

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