Plasma-Based Surface Modification of Polystyrene Microtiter Plates for Covalent Immobilization of Biomolecules

Abstract

In recent years, polymer surfaces have become increasingly popular for biomolecule attachment because of their relatively low cost and desirable bulk physicochemical characteristics. However, the chemical inertness of some polymer surfaces poses an obstacle to more expansive implementation of polymer materials in bioanalytical applications. We describe use of argon plasma to generate reactive hydroxyl moieties at the surface of polystyrene microtiter plates. The plates are then selectively functionalized with silanes and cross-linkers suitable for the covalent immobilization of biomolecules. This plasma-based method for microtiter plate functionalization was evaluated after each step by X-ray photoelectron spectroscopy, water contact angle analysis, atomic force microscopy, and bioimmobilization efficacy. We further demonstrate that the plasma treatment followed by silane derivatization supports direct, covalent immobilization of biomolecules on microtiter plates and thus overcomes challenging issues typically associated with simple physisorption. Importantly, biomolecules covalently immobilized onto microtiter plates using this plasma-based method retained functionality and demonstrated attachment efficiency comparable to commercial preactivated microtiter plates.

Gold nanoparticles assisted characterization of amine functionalized polystyrene multiwell plate and glass slide surfaces

“…This could be due to the decrease of the hydrophobicity of PS plate surfaces by the amine modification, which facilitates the interaction of water molecules with amine group of the modified surfaces through hydrogen bonds. It proves that the PS plate surfaces were modified with the amine group (North et al 2010). Similarly, Fig.…”

“…Immobilization of biomolecules on the solid surface was the key step in the diagnosis of infectious and genetic diseases, analysis of gene expression, forensic science, toxicology, drug discovery, biosensing, and environmental monitoring (Kannoujia et al 2010;North et al 2010). In general, polystyrene (PS), polypropylene, polycarbonate, and glass slide (GS) have been used as a solid materials for the immobilization of biomolecules (DNA and protein) because of their versatile chemical, physical, and surface adherence properties.…”

“…In general, polystyrene (PS), polypropylene, polycarbonate, and glass slide (GS) have been used as a solid materials for the immobilization of biomolecules (DNA and protein) because of their versatile chemical, physical, and surface adherence properties. Among these, PS and GS are most commonly used solid materials because they readily adsorb to DNA and proteins (antibodies or antigens) via non-covalent interactions (Chu et al 2005), excellent optical as well as mechanical properties, and cost-effective (North et al 2010). However, the non-covalent interactions of DNA and proteins on their surfaces also have limitations like desorption during the time of washing, poor recognizing properties, protein denaturation, loss of bioactivity, and low sensitivity (Rebeski et al 1999;Butler 2000).…”

“…The presence of functional groups and chemical status of elements, rough and smooth surface, hydrophobic and hydrophilic properties of materials was characterized by using X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and water contact angle (WCA) measurements (North et al 2010;Qin et al 2007;Idage and Badrinarayanan 1998). However, these characterization methods are cost-effective, time consuming and they are not affordable for routine modification process in the view of commercialization.…”

Antimicrobial Modifications of Polymers

“…This method demands low pressure (0.1 -100 Pa) and the presence of a working gas (usually N 2 , O 2 or Ar, CF 4 ). Plasma modifications to polymer surfaces are characterized by their weak stability over time, as polymer surfaces tend to return to their original chemical state [30][31][32][33][34][35][36][37][38][39][40].…”