Proteins detection, using

In addition to protein detection using specific antibody/antigen and aptamer/protein interactions, array detection was demonstrated based on nonspecific interactions between the CPE/dye-labeled ssDNA complexes and proteins [103]. The design concept is motivated by the fact that external agents can effectively perturb the electron coupling of optical units within the complex of CPE/ssDNA-C and in turn vary the FRET-induced fluorescence of both CPE (donor) and C (acceptor). Owing to discrepancies in local hydrophobic and charged domains of different... 

Adler M, Niemeyer CM. Enhanced protein detection using real-time immuno-PCR. In Broude NE, editor. DNA Amplification—Current Technologies and Applications. Norfolk, UK Horizon Bioscience, 2004 293-312. 

Fredriksson S, Gullberg M, Jarvius J, Olsson C, Pietras K, Gustafsdottir SM, Ostman A, Landegren U, Protein detection using proximity-dependent DNA ligation assays, Nat. Biotechnol., 20 413-411, 2002. 

Fig. 3 Protein detection using immobilized nucleic acids with tethered intercalators. Fabrication of DNA-modified gold electrodes for electrochemical analysis of protein binding and reaction (Reprinted with permission from [174], Copyright(2002) Nature Publishing Group http //www.nature.com/nbt/)...

The same authors also examined the S-nitrosylation of proteins by nNOS, in neuronal tissue from mice (Figure 11.10). Conversion of nitrosylated to biotinylated cysteines was done as above. Biotinylation was, in this case, used to extract the (formerly) nitrosylated proteins from the total mixture of cellular proteins by selective binding to solid-phase attached streptavidin. After retrieving the bound material by reduction with excess free thiol, the samples were run on a gel, blotted, and individual proteins detected using an-... 

Ochsenkuehn MA, Campbell CJ (2010) Probing biomolecular interactions using surface enhanced raman spectroscopy label-free protein detection using a g-quadmplex dna aptamer. Chem Comm 16 2799-2801... 

A typical sensorgram for protein detection using a ring resonator is shown in Fig. 5. The sensing surface is first immobilized with receptor peptide-displayed... 

Fredriksson, S., Gullberg, M., Jarvius, J., Olsson, C., Pietras, K., Gustafsdottir, S. M., Ostman, A., Landegren, U. (2002). Protein detection using proximity-dependent DNA ligation assays. Nat Biotechnol 20, 473-477. 

Protein Detection Using DNA-Modified Gold Nanoparticles, 420... 

Figure 12.13 A DNA/Au nanopaiticle-based protein detection scheme, (a) Preparation of hapten-modified nanopaiticle probes, (b) Protein detection using protein binding probes. Notice that there are nine GC pairs in sequence A, and there are only two GC pairs in sequence B. (Reproduced with permission from J.-M. Nam et al., J. Am. Chem. Soc. 2002, 124, 3820-3821. Copyright 2002 American Chemical Society.)...

Figure 4 Sensitive protein detection using the RCA (rolling circle amplification) immunoassay chip and solid-phase direct fluorescence detection. The chip is divided into 16 teflon wells, each containing an array of 256 antibodies as probes. When a protein, represented by the blue square, is captured by one of the probes, it can be recognized using a second, biotinylated antibody (red), which is subsequentiy detected by a tertiary universal antibody connected to a circular oligonucleotide. A strand-displacing DMA polymerase can use this circular template, generating a long concatemer containing many fluorescent labels.

The lowest amount of 14-3-3 protein detected using the ELISA was a concentration of 2.1 ngml Comparison of the new ELISA method with the Western blot immunoassay using samples from 63 definite or probable CJD patients and 84 samples from patients with other neurological defects resulted in a sensitivity of 92.7% for the ELISA relating to the definite CJD patients with reference to the definite and probable CJD patients, sensitivity was 88.9%. In contrast, the sensitivity of the Western blot immunoassay within the group of definite CJD patients was 95.1%, and in the group of definite and probable CJD patients it was 93.7%. The specificity for both tests was 97.6%. 

Multiplexed protein detection using the above approaches has also been developed [38]. One approach is to use bar code labeling secondary antibodies with distinct nanoparticles with easily detectable electrochemical characteristics, e.g. different dissolvable metals or quantum dots (Qdots) that can be dissolved to give ions with different reduction potentials. 

Das, J., Kelley, S.O. Protein detection using arrayed microsensor chips tuning sensor footprint to achieve ultrasensitive readout of CA-125 in serum and whole blood. Anal. Chem. 83, 1167-1172 (2011)... 

Reprinted (adapted) with permission from L. Ma, C. Wang, Y. Hong, M. Zhang, M. Su, Thermally addressed immunosorbent assay for multiplexed protein detections using phase change nanoparticles, Anal5dical Chemistry 82 (2010) 1186-1190. Copyright 2010 American Chemical Society. 

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