Proteins immunogenicity

Utility. Insufficient data Is available on the measurement of 1,25(0H)2D3 for evaluation of Its utility In clinical medicine. A major breakthrough In methodology will be needed before routine application will be possible. This could come with the development of a battery of radioimmunoassays for the measurement of all of the vitamin D metabolites. So far, however, the development of antibodies to vitamin D and Its metabolites has been limited by apparently Irreversable changes In the Important B ring of the sterol which occur during Its conjugation to Immunogenic proteins. 

Hilton, J., Dearman, R.J., Basketter, D.A. and Kimber, I., Serological responses induced in mice by immunogenic proteins and by protein respiratory allergens. Toxicol. Lett., 73,43,1994. 

The advent of immunoproteomics made possible the identification of highly immunogenic proteins that can be used for vaccine development. Proteins that have the greatest potential for eliciting a protective immune response are collectively referred to as the pathogen s immunome. Immunoproteomics has been utilized to characterize the immu-nome of B. anthracis for the development of a safer and equally efficacious vaccine. The immunoreactive proteins are first identified by using 2DE Western blot analysis in conjunction with mass spectrometry. In B. anthracis, for example, antisera from humans post-infected with anthrax were used to probe Western blots of its various... 

Figure 12.2. Identification of immunogenic proteins by 2DE/MALDI TOF-MS and Western blots probed with antisera from immunized animals or humans. (Reprinted from Khan, A. S. et al., 2006. Proteomics and Bioinformatics Strategies to Design Countermeasures Against Infectious Threat Agents. J. Chem. Inf. Model. 46 111-115. With permission.) (See color insert.)...

X. Peng, X. Ye, and S. Wang. Identification of Novel Immunogenic Proteins of Shigella Flexneri 2a by Proteomic Methodologies. Vaccine, 22(2004) 2750-2756. 

The PAP method was pioneered by Sternberger in 1979 (1). The method uses an immunological sandwich amplification and the enzyme peroxidase to effect a signal. The unique feature of this procedure is the enzyme/antibody solution, the PAP immune complex. The horseradish peroxidase enzyme, itself an immunogenic protein, is used to inoculate a given species, and a polyclonal immune response is generated against the enzyme. This antiserum is harvested and placed in solution with the enzyme so that immune complexes form that... 

Subunit vaccines Immunogenic proteins or peptide antigens customized to specific antigenic determinants are purified from tissue culture HIV, rabies virus, influenza virus, hepatitis B virus... 

It appears that fluorescence techniques are poised to receive more serious consideration for accelerated development efforts. Key obstacles remaining include stability of receptors and fluorophores, challenges that will possibly be met partially by results of the intense efforts of molecular biology, polymer science, and nanotechnology. Advances in nanomaterials such as quantum dots will likely enable improvements in optical stability and choice of excitation/emission wavelengths for various transduction methods. Stabilization of natural and artificial enzymes and rendering immunogenic protein receptors stealthy may also aid the pursuit. 

AP Teixeira-Gomes, A Cloeckaert, G Bezard, RA Bowden, G Dubray, MS Zygmunt. Identification and characterization of Brucella ovis immunogenic proteins using two-dimensional electrophoresis and immunoblotting. Electrophoresis 18 1491-1497,... 

De Groot AS, Martin W (2009) Reducing risk, improving outcomes bioengineering less immunogenic protein therapeutics. Clin Immunol 131 189-201... 

A subunit vaccine consists of one or more immunogenic epitopes, proteins, or other components of a pathogenic organism. Immunogenic epitopes can be chemically synthesized and are known as peptide vaccines, e.g., peptide vaccine candidates for foot-and-mouth disease virus. The pathogen could be disrupted, and one or more immunogenic proteins such as bacterial cell wall proteins flagella or pili and viral envelope, capsid, or nucleoproteins can be purified. The isolation of such components in purified form is sometimes cumbersome and expensive. However, bacterial exotoxins can be easily purified, inactivated, and used as toxoid vaccines. 

Bound fraction Antigen/competitor present as a complex, attached to the corresponding antibody the fi-action of the reaction mixture which contains the antigen-antibody complex Carrier protein Large non-immunogenic protein that, when coupled to a hapten, induces production of anti-hapten antibodies in an animal Competitive assay Immunoassay based on the principle of competition between the analyte in the sample ( unknown ) and competitor Competitor Antigen derivative that is modified (i.e., labelled, coupled to a large protein) and serves for indirect detection of the analsde in competitive immunoassays... 

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