Application of oriented and random antibody immobilization methods in immunosensor design

作者：Baniukevic Julija; Kirlyte Justina; Ramanavicius Arunas; Ramanaviciene Almira^*

来源：Sensors and Actuators B: Chemical , 2013, 189: 217-223.

DOI：10.1016/j.snb.2013.03.126

• SPR
• Immunosensor
• Antibody immobilization
• BLV

An immunosensor for the determination of bovine leukemia virus (BLV) antigen (gp51) using surface plasmon resonance (SPR) equipment and SPR-chip modified by oriented and randomly immobilized antibodies (anti-gp51) was developed. Different conditions of anti-gp51 reduction by 2-mercaptoethanol (2-ME) and dithiothreitol (DTT) solutions were compared and explored. The best gp51 detection sensitivity was monitored using SPR-chip modified with oriented antibody fragments (frag-anti-gp51) obtained after reduction with DTT at 50 mM concentration. The SPR sensor could detect gp51 antigenin a range from 0.01 to 0.5 mg/mL. The developed SPR sensor offered the limit of detection (LOD) as low as 0.0028 mg/mL, while the limit of quantification was as low as 0.0092 mg/mL with very good repeatability during the three detection-regeneration cycles (1-4%). Oriented immobilization of frag-anti-gp51 was found to be more suitable for the design of SPR-immunosensor for the detection of gp51 if compared with random immobilization of anti-gp51. This method was successfully applied for the detection of gp51 in blood serum samples in a rapid, reliable and selective manner.

• 出版日期2013-12