Protein Immobilization Strategies for Protein Biochips

Abstract

In the past few years, protein biochips have emerged as promising proteomic and diagnostic tools for obtaining information about protein functions and interactions. Important technological innovations have been made. However, considerable development is still required, especially regarding protein immobilization, in order to fully realize the potential of protein biochips. In fact, protein immobilization is the key to the success of microarray technology. Proteins need to be immobilized onto surfaces with high density in order to allow the usage of small amount of sample solution. Nonspecific protein adsorption needs to be avoided or at least minimized in order to improve detection performances. Moreover, full retention of protein conformation and activity is a challenging task to be accomplished. Although a large number of review papers on protein biochips have been published in recent years, few have focused on protein immobilization technology. In this review, current protein immobilization strategies, including physical, covalent, and bioaffinity immobilization for the fabrication of protein biochips, are described. Particular consideration has been given to oriented immobilization, also referred to as site-specific immobilization, which is believed will improve homogeneous surface covering and accessibility of the active site.

Inorganic nanomaterial-based biocatalysts

“…These types of compounds deprotonate upon self-assembled monolayers (SAMs), a process that neutralizes the negatively charged sulfur-containing compound assuming the formation of gold (4,5). SAMs of alkane thiols http://bmbreports.org on gold containing a functional terminal group have served to associate additional monolayers of enzymes by covalent conjugation, supramolecular association, physical adsorption and electrostatic interactions (6)(7)(8)(9)(10)(11). Previous study found that immobilization of enzyme onto gold nanoparticle's surfaces, without any surface modification, provided colloidal stability, which result to significantly enhanced catalytic activity (11).…”

Peptide-directed co-assembly of nanoprobes on multimaterial patterned solid surfaces

“…At the same time, a lack of individual Cys and His amino acids eliminates the obvious well known metal-binding mechanisms by which peptides can attach non-specifically to metal surfaces. 7,8,11,25,44,45 In QBP1, on the other hand, hydrophobic amino acids (P) have been reported to be an important constituent for silicabinding peptide sequences 30 and are probably associated with silica binding. By contrast, the present aromatic amino acids (W, Y) may contribute to the observed minor gold-binding characteristics of QBP1.…”

Serum tumor marker detection on PEGylated lipid membrane using biosensor based on total internal reflection imaging ellipsometry

“…Furthermore, as an improvement, protein A is also used for antibody immobilization on the membrane, which ensures that the binding site of the immobilized antibody remains accessible for antigen binding [37]. Table 2 is the comparison of two antibody immobilization methods.…”