Antibodies, generation

In 15-50% of cases, IgE-mediated anaphylaxis to a NMBA has been reported at the first known contact with a NMBA [1,3, 14]. This suggests a possible cross-reaction with IgE antibodies generated by previous contact with apparently unrelated chemicals drugs, such as pholcodine, cosmetics, disinfectants and industrial materials [14,... 

The EMSIL method may prove useful in cytochemical applications where lack of substrate immunogenicity precludes antibody generation, or for which enzymes specific to the... 

An alternative pharmacotherapy for cocaine dependence currently under investigation is use of a cocaine vaccine to blunt the reinforcing effects of cocaine.51 60 The basis of this pharmacotherapy is to decrease the rate of entry of cocaine into the CNS (and therefore the onset of action), by either binding cocaine with antibody generated by active immunization with a stable cocaine conjugate or by using an enzymatically active antibody specific for cocaine. 

TES-45 and TES-55 are two glycoproteins that have yet to be identified at a genetic level, but evidence has been obtained that they may also be lectins. Carbohydrate affinity chromatography with mannose-agarose shows that TES-32 selectively binds as expected, but that TES-45 is also present in small amounts (Loukas et al., 1999) unlike TES-32, TES-45 does not bind to A -ace Lylgalac t< isamine. No sequence information has yet been obtained on TES-45, but it is recognized by polyclonal antibodies generated to TES-32,... 

If a monoclonal antibody was generated by immunization with a full-length native protein rather than a peptide, then the immunized mouse will generate antibodies that recognize both linear and conformationally dependent epitopes. Only a small subset of these monoclonal antibodies will likely be useful for clinical use on formalin-fixed, paraffin-embedded tissue (FFPE) samples. Those that are useful tend to have epitopes that are linear the epitopes are not dependent on the protein s three-dimensional conformation (see Chapter 16). Therefore, for antibodies generated in response to immunization with full-length proteins, the peptides that serve as positive controls will be linear stretches of amino acids derived from the native protein sequence, as listed in protein databases. 

Cross-reactivity occurs when antigens share epitopes such that an antibody generated against one antigen will also recognize another. 

The antibodies generated in this programme were initially found to accept a broad range of substrates including acetone, fluoroacetone, 2-butanone, 3-pentanone and dihydroxyacetone. The list has now been expanded to include... 

A mechanistic analysis of antibody DF8-D5 showed it to cleave p-nitrophenyl carbamate [134] with kcat = 0.3 s l, Km = 120 p,M, and cat/ uncat = 300 at 14°C (Appendix entry 4.3) (Wentworth et al., 1997). This is some tenfold more active than a carbamatase antibody generated by Schultz... 

Figure 10.3 Whole-mass analysis of a monoclonal antibody. (A) Direct infusion of the antibody generates an envelope of high m/z ions ranging from 2000 to 3500. Deconvolution of the ion current signal gives the mass of the complete native molecule (147, 100.97 Da) and resolves some heterogeneity linked to the A-glycan structures. The major forms are consistent with molecules carrying biantennary structures capped with 0, 1, or 2 hexose (G = galactose) residues. (Data generated on an ESI-Q-Star instrument, Sciex-Applied Biosystems.)...

For immunoaffinity matrix, to affinity-purify the antibodies generated... 

Catalytic Antibodies Generated against Transition State Analogues 325... 

As noted above, the specificity of the antibody-antigen reaction is critical for obtaining reliable, interpretable results. For this reason, the antibody has to be tested rigorously, and essential controls for antibody specificity should be included in any experimental design. A comprehensive discussion on antibody generation, specificity, and testing for immunocytochemical applications can be found in references (27-29) and, for specific applications, see Chapters 17, 50, and 51. 

Nonspecific binding blocking solution 10% normal serum of the secondary antibody generating species in PBS. 

Fig. 1. Diagram illustrating the molecular interactions of the PAP procedure. The PAP complex is comprised of horseradish peroxidase bound to an antiperoxidase antibody generated in the same animal species as the primary antibody which recognized the tissue antigen of interest. The primary antibody and the PAP complex are linked via a secondary antibody generated in a second animal species against immunoglobulin of the primary animal species (A, immunoglobulin , peroxidase enzyme).

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