研究目的
To develop an immune biosensor test system for express detection of polyamines in cells of breast cancer for early diagnostics.
研究成果
The SPR-based immune biosensor can detect polyamines (spermine and spermidine) with high sensitivity, down to 0.05 ng/mL for spermidine using competitive analysis, which is suitable for early breast cancer diagnostics. The method is faster (20-40 min) and potentially cheaper than traditional ELISA. Future research should address cross-reactivity issues and apply the method to biological fluids and tissues.
研究不足
There is a 20% possibility of cross-reaction between spermidine and spermine, which could cause difficulties in analyzing biological fluids like blood plasma and urine. The method requires preliminary surface modification, which adds time, though it can be optimized.
1：Experimental Design and Method Selection:
The study used an SPR-based immune biosensor to detect polyamines (spermine and spermidine) through antigen-antibody reactions in real time. Two algorithms were employed: direct and competitive analysis.
2：Sample Selection and Data Sources:
Polyamine solutions were prepared from stock solutions with concentrations ranging from 10 to 100 ng/mL. Specific serums (antibodies) against polyamines were obtained from Abcam.
3：List of Experimental Equipment and Materials:
Materials included specific serums (ab 7318 and ab 26975 from Abcam), polyamines (spermine and spermidine from Sigma-Aldrich), and the immunobiosensor "Plazmontest" (developed at the Glushkov Institute of Cybernetics of NAS of Ukraine). Chemicals used were polyallilamine hydrochloride (PAA), protein A from Staphylococcus aureus, bovine serum albumin (BSA), and glutaraldehyde.
4：Experimental Procedures and Operational Workflow:
The transducer surface was prepared by coating with PAA, protein A, and BSA. For direct analysis, antibodies were immobilized, and polyamine solutions were applied. For competitive analysis, polyamines were conjugated with BSA and immobilized, then free polyamines and antibodies were added. Changes in resonance angle were measured after 10-20 min incubations at 25°C, with washing steps using saline solution. Calibration curves were created.
5：Data Analysis Methods:
Sensitivity and linear range were determined from calibration curves. Statistical analysis was implied from the rise in probability with concentration.
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