Silane-modified surfaces in specific antibody-mediated cell recognition

Abstract

Abstract:The immobilization of antibodies on various surfaces has been the subject of advanced research in various immunoassay-based diagnostic devices. The physical and chemical stabilities of the immobilized antibodies on a solid surface still cause many problems because upon immobilizing antibody molecules, the antigen-binding ability usually decreases. The silanization of surfaces with organosilanes carrying chemically active groups such as (3-aminopropyl)triethoxysilane (APTES) can accommodate these antigen-binding molecules in an appropriate orientation so that their functionality and binding activity are essentially retained. In this study, n-butyltrimethoxysilane (BMS) and 3-(octafluoropentyloxy)-propyltriethoxysilane (OFPOS) were used as "blocking silanes". The aims of this study were to compare the effectiveness of specific antibody binding of APTES, APTES + BMS and APTES + OFPOS and to characterize the modified surfaces by contact angle measurements and immunofluorescence measurements prior to and after immobilizing proteins. Additionally, we have evaluated the functionality of the immobilized antibodies by their abilities to bind EpCAM-positive human colon adenocarcinoma cell line (LoVo) and EpCAM-negative mouse embryonic fibroblast cell line (3T3). Cell enumeration was conducted on the basis of DAPI-positive signals and recorded using a confocal laser scanning biological microscope. The results of our study showed that the immobilization capability and reactivity of AP-TES, APTES + BMS and APTES + OFPOS differ. The modification of APTES with unreactive silanes (BMS, OFPOS) is recommended to improve the antibody binding efficiency. However, using OFPOS resulted in more effective antibody and cell binding, and it appears to be the most useful compound in specific antibody-mediated cell recognition.

Cytokeratins Biosensing Using Tilted Fiber Gratings

“…After that, TFBGs were gently rinsed using PBS and dried at RT. Our protocol was adapted from [ 26 ] and [ 27 ].…”

Controlled Engineering of Oxide Surfaces for Bioelectronics Applications Using Organic Mixed Monolayers

“…The optimization of the interface between biological objects (e.g., cells) and inorganic electronics today plays a major role in biomaterial and bioelectronics research. For example, research on biotechnical applications is interested in (i) cell adhesion to rigid or flexible substrates, − (ii) the controlled immobilization of neurons or guidance of neurite outgrowths on the surface of an electronic sensor device, − and (iii) a well-defined and stable protein pattern with areas modified with protein-resistant functionalities or protein active groups . One possible solution for these issues (listed above) is a well-controlled method to tailor the surface properties of substrates via an optimized organic interface between the substrate and the targeted biomaterial.…”

Positive influence of aminosilanes on anti-EpCAM antibody immobilization on a glass surface

“…This can be explained by the fact that a higher concentration of antibodies can affect the packing density. It may also lead to the steric hindrance in antigen binding [24][25][26]. The length and flexibility of amino-functional chains and the number of amino groups are crucial for the effective immobilization of antibodies.…”

“…The length and flexibility of amino-functional chains and the number of amino groups are crucial for the effective immobilization of antibodies. Equally, the physical and chemical stability of the immobilized antibodies still causes many problems because their binding activity can decrease after immobilization due to their random orientation [24]. That is why an appropriate immobilization method is vital to maintain the highest functionality of antibodies by site-specific orientation.…”