This review focuses on the application of novel technologies for generating biocompatible surfaces for high-throughput screening (HTS) of proteins. Various methods of coupling and spotting proteins on self-assembled monolayer (SAM) surfaces will be described along with the protein chip challenges pertaining to spot homogeneity, morphology, biocompatibility and reproducibility.

We report a novel high-throughput (HTP) protein chip platform, constructed on gold using self-assembly techniques, for conducting high quality antigen-antibody interactions. Biotinylated monolayers were used to immobilize a streptavidin surface with high packing density. This biocompatible platform was then used for detection of serum IgM antibodies. Serum samples of patients suspected to suffer from Lyme borreliosis were used to validate the protein chip platform using biotinylated peptide AAOspC8 molecules as the test probes. Various experimental parameters such as the effect of concentration of probes, targets, temperature of incubation, and their effect on the resulting signal-to-noise ratio are described in detail. Highly specific protein interaction data with a high signal-to-noise ratio were obtained with serum sample solutions as low as 1 microL/spot (1/10 diluted).

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We have developed a genetic barcode module, based on a parallel sorting facility of single nucleotide polymorphism for secure individual identification of cattle. Biotinylated allelespecific oligonucleotides were immobilized onto the predefined spots of streptavidin tethered self-assembled monolayers with long chain alkanethiols on biochips. The target DNAs for hybridization and subsequent on-chip minisequencing were produced by multiplex PCR method. After enzymatic extension, only the moiety-modified dideoxynucleotide triphosphate, when coupled to its complementary target sequence, could be detected by the corresponding antibody to the moiety in a specific and sensitive manner. The database SNPZoo was developed for storage of the sequence information consisting of cytosine/thymidine patterns. This SNP chip system can further be used in the detection of any replaceable point mutations occurring in the human and animal genes.