Cell–Surface Interactions

Livingston, Megan; Kasper, F. Kurtis

doi:10.1007/978-3-319-74854-2_7

Cited by 1 publication

(1 citation statement)

References 119 publications

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“…Nonspecific interactions of cells to a surface can be caused by van der Waals, electrostatic, and hydrophobic interactions between the cell surface molecules (e.g., lipids, proteins, and carbohydrates) and the molecules on the substrate. − Although we have used a number of strategies such as using biopolymer dextran, blocking the surface with 1% BSA solution before adding cells, blocking Fc receptors on the cells with fetal bovine serum (FBS), − using a cell culture with high (98%) viability, and using monoclonal Abs instead of polyclonal to eliminate nonspecific binding on the surface, nonspecific cell attachment on both surfaces (i.e., site-selective and passive adsorption) still occurs. This might be first due to a high density of Ab on the surface.…”

Section: Resultsmentioning

confidence: 99%

Surface Modification of Polystyrene with Boronic Acid for Immunoaffinity-Based Cell Enrichment

Shirani,

Razmjou,

Asadnia

et al. 2024

Langmuir

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Obtaining an enriched and phenotypically pure cell population from heterogeneous cell mixtures is important for diagnostics and biosensing. Existing techniques such as fluorescentactivated cell sorting (FACS) and magnetic-activated cell sorting (MACS) require preincubation with antibodies (Ab) and specialized equipment. Cell immunopanning removes the need for preincubation and can be done with no specialized equipment. The majority of the available antibody-mediated analyte capture techniques require a modification to the Abs for binding. In this work, no antibody modification is used because we take advantage of the carbohydrate chain in the Fc region of Ab. We use boronic acid as a cross-linker to bind the Ab to a modified surface. The process allows for functional orientation and cleavable binding of the Ab. In this study, we created an immunoaffinity matrix on polystyrene (PS), an inexpensive and ubiquitous plastic. We observed a 37% increase in Ab binding compared with that of a passive adsorption approach. The method also displayed a more consistent antibody binding with 17 times less variation in Ab loading among replicates than did the passive adsorption approach. Surface topography analysis revealed that a dextran coating reduced nonspecific antibody binding. Elemental analysis (XPS) was used to characterize the surface at different stages and showed that APBA molecules can bind upside-down on the surface. While upside-down antibodies likely remain functional, their elution behavior might differ from those bound in the desired way. Cell capture experiments show that the new surface has 43% better selectivity and 2.4-fold higher capture efficiency compared to a control surface of passively adsorbed Abs. This specific surface chemistry modification will allow the targeted capture of cells or analytes with the option of chemical detachment for further research and characterization.

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