Immunochemical blocking

Baldo BA, Fisher MM, Pham NH On the origin and specificity of antibodies to neuromuscular blocking (muscle relaxant) drugs an immunochemical perspective. Clin Exp Allergy 2009 39 325. 

Habeeb, A.F.S.A., and Atassi, M.Z. (1970) Enzymatic and immunochemical properties of lysozyme. Evaluation of several amino group reversible blocking reagents. Biochemistry 9, 4959—4944. 

Positive results from the in vitro micronucleus test indicate that the test substance induces chromosome damage and/or damage to the cell division apparams, in cultured mammahan somatic cells. Immunochemical labehng (FISH fluorescence in sim hybridization) of kinetochores, or hybridization with general or chromosome specific centromeric/telomeric probes can provide useful information on the mechanism of micronucleus formation. Use of cytokinesis block facilitates the acquisition of the additional mechanistic information (e.g., chromosome nondisjunction) that can be obtained by FISH techniques. The micronucleus assay has a number of advantages over metaphase analysis performed to measure chromosome aberrations (see OECD TG 487 draft). 

The first step in immunochemical detection of proteins after electrotransfer is blocking the support with an inert material to inactivate further non-specific binding of protein. The blocking reagent should cover the membranes at those areas where no blotted protein is bound and should not react with any of the reactants of immunochemical detection cascade as indicated by no non-specific staining, i.e., resulting in blank background of the membrane. 

The polysaccharide from Hansenula polymorpha 52-251 has a block type of structure of the main chain, with (1— -2)- and (1— 6)-linked a-D-mannopyranosyl units. The side chains also contain (l- 2) links interspersed with (1— 6) links. Immunochemical experiments revealed the presence of a-D-glucopyranosyl phosphate determinants,118 in common with the polysaccharide of Hansenula wingei, a yeast of interest because of its sexual, agglutination factors.104... 

Electroblots from SDS-PAGE [9] in 12% gels were stained either for total protein [9], glycoprotein [1] or immunochemically using Tween blocking, primary antiserum diluted 1 1000 (rabbit) or 1 10 (culture supernatant) and peroxidase-labelled second antibody. 

Fig. 7.10 Concentration-dependent inhibition of binding of IgE antibodies specific for tertiary and quaternary methyl (mainly) and ethyl eunino groups by neuromuscular blocking drugs. Side-by-side quantitative comparisons of inhibitory potencies of rocuronium and alcuronium with six other neuromuscular blocking drugs. Refer to Table 7.11 for the precise quantitative relationships. The dashed line indicates 50 % inhibition. From Pham NH et al. Studies on the mechanism of multiple drug allergies. Structural basis of drug recognition. J Immunoassay Immunochem. 2001 22 47. Reprinted with permission from Taylor Francis...

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