Author:
Jena Subas Chandra,Shrivastava Sameer,Saxena Sonal,Kumar Naveen,Maiti Swapan Kumar,Mishra Bishnu Prasad,Singh Raj Kumar
Abstract
Abstract
We report detection of Baculoviral inhibitor of apoptosis repeat containing-5 (BIRC5) protein biomarker in dog serum by label-free surface plasmon resonance (SPR) immunosensor. Initially, overexpression of BIRC5 in canine mammary tumour (CMT) tissues was confirmed by real-time PCR. Recombinant BIRC5 was produced and protein specific antibodies developed in guinea pig specifically reacted with native protein in immunohistochemistry and immunocytochemistry. SPR immunosensor was developed by fabricating anti-BIRC5 antibodies on gold sensor disc. The equilibrium dissociation constant, (KD = kd/ka) was 12.1 × 10−12 M; which indicates that antibodies are of high affinity with sensitivity in picomolar range. The SPR assay could detect as low as 6.25 pg/ml of BIRC5 protein in a calibration experiment (r2 = 0.9964). On testing real clinical samples, 95% specificity and 73.33% sensitivity were recorded. The average amount of serum BIRC5 in dogs with CMT was 110.02 ± 9.77 pg/ml; whereas, in non-cancerous disease conditions, 44.79 ± 4.28 pg/ml and in healthy dog sera 30.28 ± 2.99 pg/ml protein was detected. The SPR immunosensor for detection of BIRC5 in dog sera is reported for the first time and this may find prognostic and diagnostic applications in management of CMT. In future, ‘on-site’ sensors can be developed using this technique for near-patient testing.
Funder
Department of Biotechnology, Ministry of Science and Technology
Indian Council of Agricultural Research
Publisher
Springer Science and Business Media LLC
Reference59 articles.
1. Si, C. Y. Rapid detection of Escherichia coli O157: H7 using surface plasmon resonance (SPR) biosensor. Spectrosc. Spectral Anal. 31(10), 2598–2601 (2011).
2. Piliarik, M., Parova, L. & Homola, J. High-throughput SPR sensor for food safety. Biosens. Bioelectron. 24(5), 1399–404 (2009).
3. Rasooly, A. Surface plasmon resonance analysis of staphylococcal enterotoxin B in food. J. Food Prot. 64(1), 37–43 (2001).
4. Hodnik, V. & Anderluh, G. Toxin detection by surface plasmon resonance. Sensors (Basel) 9(3), 1339–54 (2009).
5. Gupta, G. et al. Surface plasmon resonance detection of biological warfare agent Staphylococcal enterotoxin B using high affinity monoclonal antibody. Thin Solid Films. 519(3), 1171–1177 (2010).
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