Antibody-based biosensor to detect oncogenic splicing factor Sam68 for the diagnosis of lung cancer
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ORIGINAL RESEARCH PAPER
Antibody-based biosensor to detect oncogenic splicing factor Sam68 for the diagnosis of lung cancer B. Sumithra . V. S. P. K. Sankara Aditya Jayanthi . Hari Chandana Manne . Rashmika Gunda . Urmila Saxena . Asim Bikas Das
Received: 24 February 2020 / Accepted: 28 June 2020 Ó Springer Nature B.V. 2020
Abstract Objective The present work aimed to investigate the potential utility of Sam68 protein as a prognostic marker in lung cancer. Then an electrochemical immunosensor is fabricated that is sufficiently sensitive to detect Sam68. Results Analysis of stage-specific Lung cancer microarray data shows that differential expression of Sam68 is associated with cancer stage and monotonically increases from early tumor stage to advanced metastatic stage. Moreover, the higher expression of Sam68 results in reduced survival of lung cancer patients. Based on these observations, an electrochemical immunosensor was developed for the quantification of Sam68 protein. The target protein was captured by the Anti-Sam68 antibody that was immobilized on the modified Glassy carbon electrode. The
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02951-9) contains supplementary material, which is available to authorized users. B. Sumithra V. S. P. K. S. A. Jayanthi H. C. Manne R. Gunda U. Saxena (&) A. B. Das (&) Department of Biotechnology, National Institute of Technology Warangal, Warangal, Telangana 506004, India e-mail: [email protected]; [email protected] A. B. Das e-mail: [email protected]; [email protected]
stepwise assembly process was characterized by cyclic voltammetry and electrochemical impedance spectroscopy. This fabricated immunosensor displayed good analytical performance in comparison to commercial ELISA kit with good sensitivity, lower detection limit (LOD) of 10.5 pg mL-1, and wide linear detection range from 1 to 5 lg mL-1. This method was validated with satisfactory detection of Sam68 protein in lung adenocarcinoma cell line, NCIH23. Besides, spike and recovery assay reconfirm that the sensor can precisely quantify Sam68 protein in a complex physiological sample. Conclusion We conclude Sam68 as a valuable prognostic biomarker for early detection of lung cancer. Moreover, we report the first study on the development of an electrochemical immunosensor for the detection of Sam68. The fabricated immunosensor exhibit excellent analytical performance, which can accurately predict the lung cancer patient pathological state. Keywords Splicing factor Prognostic Diagnostic Biomarker Lung cancer Biosensors
Introduction The Src-associated protein in mitosis (Sam68), also known as KHDRBS1, is a member of signal
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Biotechnol Lett
transduction and activation of RNA (STAR) family of RNA-binding proteins (Lukong and Richard 2003). Sam68 is involved in both RNA processing and signal transduction pathways due to the presence of KHdomain (K homology) and SH-domain (Src h
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