Sandwich-format ECL immunosensor based on Au star@BSA-Luminol nanocomposites for determination of human chorionic...

A sandwich-configuration electrochemiluminescence (ECL) immunosensor based on Au star@BSA-Luminol nanocomposites for ultrasensitive determination of human chorionic gonadotropin (HCG) has been developed. In this work, nanostructured Polyaniline hydrogels (Pani) decorated with Pt nanoparticles (Pani/Pt) were utilized to construct the base of this immunosensor, greatly increasing the amount of loaded capture antibodies (Ab₁) via linkage reagent glutaraldehyde (GA). The used conducting Pani/Pt nanocomposites possessed the unique features, such as large surface area, high electron transfer speed and favorable electrocatalytic activities of hydrogen peroxide, which offered a prominent platform for this sandwich-sensor and acted as efficient ECL signal amplifier also. Furthermore, we employed horseradish peroxidase (HRP) to block the nonspecific binding sites instead of commonly used bovine serum albumin (BSA), which further amplified the signal of luminol in the present of hydrogen peroxide (H₂O₂). In addition, Au star@BSA nanocomposites with excellent water solubility, low-toxicity and great biocompatibility were prepared and used to immobilize HCG detection antibodies (Ab₂) and luminescent material luminol. Then, the above-synthetized Luminol-Au star@BSA-Ab₂ complex was attached to the modified sensor by sandwiched immunoreactions. Under the optimized conditions, the proposed immunosensor exhibited a sensitive detection of HCG in a wide linear range from 0.001 to 500mIUmL^-1 with a detection limit of 0.0003mIU/mL (S/N = 3). All the results indicated that such a sandwiched HCG immunosensor exhibited favorable ECL analytical performance. This developed method may be potentially used to recognize other clinical protein and display a novel ideal to construct an immunosensor.