Protein immuno assay

Funke, C. et al., Harbor seal (Phoca vitulina) C-reactive protein (C-RP) purification, characterization of specific monoclonal antibodies and development of an immuno-assay to measure serum C-RP concentrations, Vet. Immunol. Immunopathol., 59, 151, 1997. 

Viral structural proteins expressed by baculovirus infected insect cells assemble into multimeric structures that resemble viral core-like particles and virus-like particles (CLPs and VLPs, respectively). This presentation has brought the attention of researchers for the potential use of these structures as safe immunological reagents for virus or antibody detection in enzyme immuno assays, as vaccines, and more recently, as gene delivering systems for gene therapy [9]. 

Figure 3 Adsorption IgG to a series of poly(lauryl methacrylate-co-glycerol methacylate-co-ethylene glycol dimethacrylate)s Results from immuno assay, optical density is directly proportional to amount of adsorbed protein...

Tanaka T, R. Sato R, Kamiya S, Obata K, Tamiya S, Matsunaga T. (2000). Fully-automated heterogeneous immuno assay of human insulin using protein A-bacterial magnetic particle complexes. Anal. Chem. 72,3518- 3522... 

The energy transfer rate from the donor to the acceptor deereases with the sixth power of the distanee. Therefore it is noticeable only at distanees shorter than 10 nm [308]. FRET is used as a tool of immuno-assay teehniques and of eell and tissue fluoreseenee imaging see Sect. 5.7.6, page 149. Different proteins are labelled with the donor and the acceptor, and FRET is used as an indieator of the binding state of these proteins. Distances on the nm scale ean be determined by measuring the FRET effieiency quantitatively. 

A bridge between natural and artificial macromolecular metal complexes is the interaction of metal ions/complexes with peptides/proteins [70], nucleic acids/DNA [71,72], enzymes [73], steroids [74], carbohydrates [75]. Biometal-organic chemistry concentrates on such complexes [15], The reason for the increasing interest in this field lays in medical applications of metal complexes [16,76] (cancer, photodynamic therapy of cancer -immuno-assays, fluorescence markers, enantioselective catalysis, template orientated synthesis of peptides) as exemplarily shown below. 

Doi, H., Shibata, H., Shoji, M., Sakai, S., and Akiyama, H. (2008). A reliable enzyme linked immuno-sorbent assay for the determination of walnut proteins in processed foods. /. Agric. Food Chem. 56, 7625-7630. 

There are many examples of ELIS As used for detecting host cell impurities in the literature. Pauly et al.12 developed an ELISA to detect impurities in erythropoietin that had a detection limit of around 0.05 ng/ml. SDS polyacrylamide gel and Western blot analysis were used to confirm the spectrum of proteins detected and to demonstrate the specificity of the antibody preparation. Anicetti et al.14 describe an assay for the detection of E. coli proteins in recombinant DNA-derived human growth hormone. Whitmire and Eaton15 report on an immuno-ligand assay for quantitation of process-specific E. coli host cell contaminant proteins in a recombinant bovine somatotropin. 

Ghobrial, I.A., D.T. Wong, and B.G. Sharma (1997). An immuno-ligand assay for the detection and quantitation of contaminating proteins in recombinant human erythropoietin (r-HuEPO). BioPharm-Technol Bus 10(1) 42 45. 

Recent evidence confirms that species differences can involve more than one aspect of PPARa-mediated regulation of gene expression. The insensitivity of human liver to rodent peroxisome proliferators is associated with low levels of expression of PPARa in human liver. Marked species differences in the expression of PPARa mRNA have been demonstrated between rodent and human liver, with the latter expressing 1-10% of the levels found in mouse or rat liver (Palmer et al, 1994 Tugwood et al, 1996 Palmer etal, 1998). Using a sensitive and specific immuno/DNA binding assay. Palmer et al (1998) have shown that active PPARa protein is expressed at variable concentrations in human livers. The study compared 20 different human livers and found that those with the highest levels of PPARa protein expression contained less than 10% of the level in mice. Most of the samples (13/20) contained no detectable PPARa activity, but did... 

Fig. 3. Comparison of different enzyme-linked immuno sorbent assay (ELISA) methods adapted for immuno-polymerase chain reaction (IPCR). Dependent on the purification grade of the sample to be analyzed and the availability of specific and functionalized antibodies, several typical ELISA protocols were adapted to IPCR. In the direct approach (A), the pure antigen is immobilized to the microplate surface and subsequently detected by a labeled specific antibody. If no labeled antibody is available (e.g., because of unpurified ascites fluid containing the antibody or loss in activity following labeling), a standardized labeled secondary species-specific antibody is used for detection of the primary antigen-specific antibody (B). For the detection of the antigen from matrices such as serum, plasma, tissue homogenate, and so on, a capture antibody immobilized to the microplate surface was used either in a direct (C) or indirect (D) sandwich approach, with the latter one additionally including a secondary species-specific detection antibody. For different methods of coupling antibody and DNA, abbreviated by in this figure, compare Fig. 2. Note that protein A chimeras (Fig. 2A) are not compatible with capture antibodies (Fig. 3C, D).

Adler M, Wacker R, Niemeyer CM. A real-time immuno-PCR assay for routine ultrasensitive quantification of proteins. Biochem Biophys Res Commun 2003 308(2) 240-250. 

Tsujii H, Okamoto Y, Nakano H, Watanabe K, Naraki T. Application of immuno-PCR assay to detect an ultra-low level plasma protein, pivka-II. International Hepatol Commun 1995 3(1000) 154. 

Metcalf, J.S., Bell, S.G., and Codd, G.A. 2001. Colorimetric immuno-protein phosphatase inhibition assay for specific detection of microcystins and nodularins of cyanobacteria. Appl Environ Microbiol 67 904—909. 

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