Current Advances in Antibody Immobilization on Different Surfaces and Beads

Abstract

Antibody immobilization is of considerable interest for miscellaneous fields of interest, e.g. detecting biomarkers in cancer diagnostics, e.g. prostate specific antigen (PSA) for prostate cancer and pathogens like Escherichia coli, a foodborne pathogen. For this purpose, specific antibody captures are required and all of them should guarantee highest reproducibility and capacity possible. Especially in clinical applications very complex and specific immunoassays are needed. However, the complexity of pinning antibodies necessitates particular methods concerning the immobilization technique itself-because of their structure and specificity-and of course the analyzing methods. Glass, polymers, gold and even fullerene beads have been used as carriers, treated with various spacers and activation steps within the last few years. Furthermore, considerable attention is drawn to the regeneration of matrices and their capabilities for further antibody attachment. Commonly applied verification tools for the detection of the trapped antibodies include impedance spectroscopy (IS), atomic fluorescence microscopy (AFM), electrophoresis (EP), enzyme linked immunosorbent assays (ELISA), quartz crystal microbalance (QCM) and infrared spectroscopy (IR). Matrix assisted laser desorption/ionization-time-offlight mass spectrometry (MALDI-TOF/MS) is also a useful tool for proving the success of new antibody fixing procedures. Immobilization techniques have been improved concerning their reproducibility, binding capacity and high specificity. In this review, recent developments of antibody (Ab) immobilization are summarized, the individual applications are mentioned, advantages and disadvantages are discussed in detail.

Adsorption and Unfolding of Lysozyme at a Polarized Aqueous–Organic Liquid Interface

“…A variety of methods have been used to detect the adsorption of proteins and polypeptides at interfaces, such as quartz crystal microbalance (QCM), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), electrochemistry, and surface plasmon resonance (SPR). − Protein behavior at the interface between two immiscible electrolyte solutions (ITIES) has emerged over recent years as a strategy for label-free detection and quantification at low concentrations. , Protein adsorption between two electrolyte solutions is a complex process that involves various kinds of interactions, such as van der Waals, hydrogen bonding, and electrostatic forces. Therefore, to study the process of adsorption at interfaces in atomistic detail is a very challenging experimental task.…”

Comparison of 1-Ethyl-3-(3-Dimethylaminopropyl) Carbodiimide Based Strategies to Crosslink Antibodies on Amine-Functionalized Platforms for Immunodiagnostic Applications

“…The immobilization of antibodies on the bioanalytical platforms is the most critical step in immunodiagnostics as it directly impacts their analytical performance [ 1 ]. A wide range of antibody immobilization strategies [ 2 , 3 , 4 , 5 ] are available such as physical adsorption, orientated binding by intermediate proteins, covalent binding, biotin-avidin interactions, affinity tags, and site-specific binding. However, the strategies based on the covalent binding of antibodies are the most prominent as they lead to rapid, leach-proof and highly stable antibody binding with high immobilization density.…”

Trends and Perspectives in Immunosensors

“…Other authors have used amino-silanes combined with a bi-functional N-hydroxysuccinimide (NHS) or a mercapto-silane combined with maleimido-NHS [14]. In relation to gold surfaces, these are generally functionalised using a thiol or amino bi-functional cross-linker, which will attach to the gold, leaving another reactive group free to bind the protein of interest [15].…”