[show abstract][hide abstract] ABSTRACT: Our research in immunotechnology focuses on the cells and molecules of the immune system for various biomedical applications.
Applied research as well as projects addressing fundamental issues is being pursued within four main project areas: 1) antibody
technology, 2) proteomics, 3) cancer, and 4) allergy. Within the proteomics project, we have taken advantage of the strong
background in antibody engineering to design antibody-based micro- and nanoarrays for applications ranging from focused assays
to proteome-scale analysis. Antibody-based microarray is a novel technology that holds great promise in proteomics. The microarray
can be printed with thousands of recombinant antibodies carrying the desired specificities, the biological sample added (e.g.,
an entire proteome), and virtually any specifically bound analytes detected. The microarray patterns generated can then be
converted into proteomic maps, or molecular fingerprints, revealing the composition of the proteome. Global proteome analysis
and protein expression profiling, by using this tool, will provide new opportunities for biomarker discovery, drug target
identification, and disease diagnostics and will give insight into disease biology. Ultimately, we apply this novel technology
platform within our cancer and allergy projects to perform high-throughput disease proteomics.
[show abstract][hide abstract] ABSTRACT: Lipid bilayers containing 5% nitrilotriacetic acid (NTA) lipids supported on SiO2 have been used as a template for immobilization of oligohistidine-tagged single-chained antibody fragments (scFvs) directed against cholera toxin. It was demonstrated that histidine-tagged scFvs could be equally efficiently coupled to an NTA-Ni2+-containing lipid bilayer from a purified sample as from an expression supernatant, thereby providing a coupling method that eliminates time-consuming protein prepurification steps. Irrespective of whether the coupling was made from the unpurified or purified antibody preparation, the template proved to be efficient for antigen (cholera toxin) detection, verified using quartz crystal microbalance with dissipation monitoring. In addition, via a secondary amplification step using lipid vesicles containing GM1 (the natural membrane receptor for cholera toxin), the detection limit of cholera toxin was less than 750 pM. To further strengthen the coupling of scFvs to the lipid bilayer, scFvs containing two histidine tags, instead of just one tag, were also evaluated. The increased coupling strength provided via the bivalent anchoring significantly reduced scFv displacement in complex solutions containing large amounts of histidine-containing proteins, verified via cholera toxin detection in serum.
[show abstract][hide abstract] ABSTRACT: We wish to alert your readers to MIAPAR, the minimum information about a protein affinity reagent. This is a proposal developed within the community as an important first step in formalizing standards in reporting the production and properties of protein binding reagents, such as antibodies, developed and sold for the identification and detection of specific proteins present in biological samples. It defines a checklist of required information, intended for use by producers of affinity reagents, quality-control laboratories, users and databases (Supplementary Table 1). We envision that both commercial and freely available affinity reagents, as well as published studies using these reagents, could include a MIAPAR-compliant document describing the product's properties with every available binding partner. This would enable the user or reader to make a fully informed evaluation of the validity of conclusions drawn using this reagent (Fig. 1).