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Surface plasmon resonance immunosensor for label-free detection of BIRC5 biomarker in spontaneously occurring canine mammary tumours.

We report the detection of apoptosis inhibitor Baculoviral repeat-containing 5 (BIRC5) serum protein biomarkers in dogs by surface plasmon resonance (SPR) immunosensor free label. Initially, overexpression BIRC5 in canine mammary tumors (CMT) network is confirmed by real-time PCR. BIRC5 recombinant proteins are produced and developed specific Horse Recombinant Proteins

 antibodies in guinea pig specifically react with native proteins in immunohistochemistry and immunocytochemistry. SPR immunosensor antibody developed by fabricating anti-BIRC5 the sensor disk of gold. The equilibrium dissociation constant (KD = kd / ka) is 12.1 × 10-12 M; indicating that high affinity antibodies with a sensitivity in the picomolar range. SPR test can detect as low as 6.25 pg / ml protein in experimental calibration BIRC5 (r2 = 0.9964). 

In real clinical sample testing, 95% specificity and sensitivity recorded 73.33%. The average amount BIRC5 serum in dogs with CMT was 110.02 ± 9.77 pg / ml; whereas, in a non-cancerous disease conditions, 44.79 ± 4.28 pg / ml and in sera of healthy dog ​​30.28 ± 2.99 pg / ml protein was detected. SPR immunosensor for detecting BIRC5 in dog sera are reported for the first time and it can find a prognostic and diagnostic applications in the management of CMT. In the future, 'in place' sensor can be developed using this technique for near-patient testing.
Surface plasmon resonance immunosensor for label-free detection of BIRC5 biomarker in spontaneously occurring canine mammary tumours.

Ipad-loaded N-trimethyl Chitosan Nanoparticles Can Efficiently Protect against Shigella flexneri Guinea Pigs.


Shigella flexneri is the pathogen responsible for shigellosis worldwide, especially in developing countries. Many immunogenic antigen has been introduced as a vaccine candidate against Shigella, including the N-terminal region of antigen ipad (NIpaD) .

To evaluate the efficiency of the O-metylated free trimethyl chitosan nanoparticles (NP TMC) in the oral delivery NIpaD.TMC synthesized by two-step method of chitosan high molecular weight. NIpaD recombinant protein used as the immunogen. This protein is expressed in E. coli BL21 (DE3) and characterized by gel electrophoresis. The NIpaD-loaded NP TMC synthesized by ionic gelation method and characterized by electron microscopy. 

NP is administered orally to mice and Native Recombinant Proteins specific humoral immune response and mucosal assessed by serum IgG and secretory IgA, respectively. The protective formulations rated with keratoconjunctivitis (Sereny) test.The immunized guinea pigs showed a significant rise in specific rNIpaD serum IgG and IgA titers feces. Specific secretory IgA were detected in an eye-wash. Sereny test results showed that the immunized animals vaccinated with iPad-loaded TMC NPS tolerated wild type Shigella flexneri 2a in the Sereny test
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