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Development of a highly sensitive mouse monoclonal antibody for screening ALK‐rearrangements in lung cancers
Author(s) -
Shen Yi,
Jin Mulan,
Wu Yipan,
Zhang Yungang,
Yuan Kehu,
Chu Boyang,
Qi Lili,
Liu Huibo,
Wang Chenlin,
Wang Guangli,
Shu Youmin,
McDowell Julie,
Wei Haitao,
Chen Caiwei,
Ma Donghui,
He Weiwu,
Chen Jian
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.417.11
Subject(s) - crizotinib , anaplastic lymphoma kinase , ceritinib , lung cancer , fluorescence in situ hybridization , cancer research , immunohistochemistry , biology , population , pathology , medicine , microbiology and biotechnology , gene , genetics , environmental health , malignant pleural effusion , chromosome
The anaplastic lymphoma kinase (ALK) rearrangements, mostly EML4‐ALK fusion, occur in 3‐7% of lung cancer patients and define a patient population that could respond to receptor tyrosine kinase inhibitor Crizotinib and Ceritinib. Currently, ALK testing is mostly conducted by either fluorescence in situ hybridization (FISH) or polymerase chain reaction (PCR) methods, either of which has its own limitations. The detection of the ALK protein in lung cancer patient tissues by IHC was found to be difficult mostly due to the low abundance of its fusion product. To overcome this limitation, we have developed an ALK mouse monoclonal antibody (clone 1A4) that is more sensitive than a current rabbit ALK antibody D5F3. Our initial analyses revealed that 1A4 can correctly identify all 5 EML4‐ALK positive samples that were previously confirmed by QPCR tests, while did not yield significant background on all 30 EML4‐ALK negative lung cancer samples. By using a different IHC detection system, we further tested 1A4 on 17 ALK‐positive and 10 ALK‐negative lung cancer biopsy specimens that have been validated by either FISH or PCR from another hospital. The IHC result of the antibody 100% agreed with the previous molecular diagnosis. The high concordance of the IHC results by the novel ALK antibody with other DNA/RNA based detection methods suggested that 1A4 could be used routinely for screening ALK‐rearrangement genetic events among patients. This research is partly supported by Chinese 863 research grant number 2014AA020909.