A portable surface plasmon resonance (SPR) measurement prototype integrated with a multiple protein-patterned SPR biochip is introduced for label-free and selective detection of human immunoglobulin-G (H-IgG). The polyclonal anti-H-IgG antibodies derived from goat, rabbit, and mouse were immobilized through polydimethylsiloxane (PDMS) microchannels to fabricate the patterned SPR biochip. The PDMS surface was functionalized using 3-aminopropyltrimethoxysilane and bonded to carbodiimide-activated gold substrates to construct irreversibly bonded hydrophilic microfluidic chip at room temperature. For SPR measurement, a custom-made system is developed with a high angular scanning accuracy of 0.005° and a wide scanning range of 30°-80° that avoids the conventional requirement of expensive goniometric stages and detector arrays. The SPR biochip immobilized with 750 μg/mL goat anti-H-IgG demonstrated detection of H-IgG with a detection limits of 15 μg/mL, and linear response through a wide concentration range (15-225 μg/mL) of high coefficient of determination (R2 = 0.99661). The selectivity of the sensor was investigated by exposing them to two different non-specific targets (bovine serum albumin and polyvalent antivenom). The results indicate negligible sensor response towards nonspecific targets (0.25° for 30 μg/mL bovine serum albumin (BSA) and 0.25° for 30 μg/mL polyvalent antivenom) in comparison to H-IgG (1.5° for 30 μg/mL).
Keywords: Kretschmann; human immunoglobulin-G; microfluidics; polydimethylsiloxane; surface plasmon resonance.
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