Assay strategies

An alternative assay format entails the use of virus-specific DNA probes. These can be used to screen the biopharmaceutical product for the presence of viral DNA. The assay strategy is similar to the dot blot assays used to detect host-cell-derived DNA contaminants, as discussed earlier. 

R. B. McAlister, K. Reitz, G. Kang, R. Nakatsu, T. Green, C. Library-Based Lead Compound Discovery Antioxidants by an Analogous Synthe-sis Deconvolutive Assay Strategy, J. Org. Chem. 1994, 59, 5862. (e) Moon, H.-S. Schore, N. E. Kurth, M. J. A Polymer-Supported C2-Symmetric Chiral Auxiliary Preparation of Non-Racemic 3,5-Disubstituted-y-Butyro-... 

LJ Reynolds, WN Washburn, RA Deems, EA Dennis. Assay strategies and methods for phospholipases. In EA Dennis, ed. Methods in Enzymology 197. Orlando FL Academic Press, 1991, pp 3-23. 

Kurth MJ, Ahlberg Randall LA, Chen C, Melander C, Miller R, Library-based lead compound discovery antioxidants by an analogous synthesis deconvolu-tive assay strategy, J. Org. Chem., 59 5862-5864, 1994. 

Because of the lack of adequately designed studies and standardized assays for many products the consequences of their immunogenicity is unclear. So assuming a product to be non-immunogenic based on literature is impossible. In other words, every new protein should be assumed to be immunogenic unless proved otherwise with a well-designed and validated assay strategy. 

The definition of the assay strategy and readout determines the requirements for the labeling of the substrate (see 2.3 covering assay formats). 

Figure 3 Common assay strategies for evaluating binding to a glycan array.

Fig. 2 Flow cytometric differential detergent resistance (FCDR) assay strategy for the detection of DRM association of membrane proteins. A Lipid raft-associated plasma membrane proteins show high resistance to detergent treatment. B Nonraft-assocrated proteins are rapidly dissolved by even low concentrations of nonionic detergents in a short time. These processes can be followed by measnring fluorescence means before and after 5 min of detergent treatment by flow cytometry. (Adapted from Gombos et al. 2004 Wolf et al. 2006)...

Jakubik, J., Wess, J., 1999. Use of a sandwich enzyme-linked immunosorbent assay strategy to study mechanism of G protein-coupled receptor assembly. J. Biol. Chem. 274, 1349-1358. 

This chapter gives an overview of the current knowledge about the immunogenicity of therapeutic proteins. First the mechanisms by which antibodies are formed will be discussed, followed by the factors that can influence the immune response. An assay strategy for the detection of antibodies in human sera is discussed. The... 

The assays described in this chapter are designed to quantify the relative rate with which an NLS green fluorescent protein (NLS-GFP) fusion protein moves in and out of the nucleus, in normal and mutant yeast cells, under a variety of physiological and nonphysiological conditions. The assay strategy circumvents the need to microinject substrates, which is not generally feasible in yeast. Al-... 

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