Antibody microarray

Antibody microarray

An antibody microarray is a specific form of protein microarrays, a collection of capture antibodies are spotted and fixed on a solid surface, such as glass, plastic and silicon chip for the purpose of detecting antigens. Antibody microarray is often used for detecting protein expressions from cell lysates in general research and special biomarkers from serum or urine for diagnostic applications.

Background

The theoretical background for protein microarray-based ligand binding assays was initially developed by Ekins et al. in the late 1980s. According to the model, antibody microarrays not only would permit simultaneous screening of an analyte panel, but would also be more sensitive and rapid than conventional screening methods. Interest in screening large protein sets only arose as a result of the achievements in genomics by DNA microarrays and the Human Genome Project.

The first array approaches attempted to miniaturize biochemical and immunobiological assays usually performed in 96-well microtiter plates. 96-well antibody arrays were first created with 144 elements each for standard enzyme-linked immunosorbent assays (ELISAs). Similar arrays were used to measure prostate-specific antigen (PSA) and cytokines.

Filter membranes were also initially used because of their superior protein binding capacity. They were mostly probed with antibodies using ELISA techniques. A low density array of 48 purified proteins involved in transcription was developed for the investigation of specific interactions of proteins with radiolabeled DNA, RNA, ligands, and other small chemicals. A membrane-based high density array was developed for the purpose of screening a human fetal brain cDNA expression library consisting of 37830 clones. Purified proteins were spotted onto PVDF membranes at a density of 300 samples/cm2. Other filter based arrays were constructed but the limitations were the low resolution and considerable background making it difficult to use them in applications with limiting sample quantities such as protein expression profiling of tumor biopsies.

Protein arrays are compromised of a library of proteins or antibodies immobilized in a 2D addressable grid on a chip. Protein microarray biochips extract and retain targets from liquid media and are distinct from microfluidic biochips, which separate and process proteins in a transport medium in situ using microfluidic devices. A typical array may contain 103–104 spatially distinct elements within a total area of 1 cm2. [ [http://www.molecularstation.com/protein-microarrays/#intro protein microarray chips ] ]

Related microarray technologies also include protein microarrays, DNA microarrays, tissue microarrays and chemical compound microarrays.

References


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