Covalent coupling of lysozyme to amino-polystyrene particles utilizing a heterobifunctional coupling agent, SPDP, and to aldehyde particles was studied using Particle Concentration Fluorescence Immunoassay (PCFIA). The results showed that these methods can be used to couple lysozyme covalently to amino-polystyrene and aldehyde particles.
The modification of amino-polystyrene particles with SPDP offers the advantage that the resulting pyridyldisulfide particles can be used to react specifically with the sulfhydryl group of proteins, or they can be reduced further with DTT to form sulfhydryl particles. The aldehyde particles can be used to couple covalently to proteins without any coupling agent.
The activity of the lysozyme coated particles depends upon the orientation of lysozyme on the particles. Higher activity was obtained when the amino groups of the lysozyme were utilized to couple to the particles covalently.
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