Antibodies antibody generation

The sensitivity of RlAs can be enhanced using a sandwich technique. This method uses two different monoclonal antibodies (antibodies generated by a single clone of plasma cells rather than multiple clones), each of which recognizes a different specific portion of the hormone s structure. The first antibody, attached to a solid support matrix such as a plastic culture dish, binds the hormone to be assayed. After exposure of the patient sample to this first antibody, the excess plasma is washed away, and the second antibody (which is radiolabeled) is then incubated with the first antibody-hormone complex. The amount of binding of the second (labeled) antibody to the first complex is proportional to the concentration of the hormone in the sample. 

Muronomab-CD3 is a murine monoclonal antibody directed against the CD3 complex of the T-lymphocyte antigen receptor. This drug selectively diminishes the T-lymphocyte pool resulting in a strong lymphopenia. Similar to other nonhuman antibodies the generation of human antimurine antibodies (HAMA) limits its long-term use. 

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. 

Figure 21.7 An intact A-B subunit toxin molecule may be activated with 2-iminothiolane with good retention of cytotoxic activity. The thiolated toxin then may be conjugated with SPDP-activated antibody to generate the immunotoxin conjugate through a disulfide bond.

I immediate Soluble Clonal expansion B cells Cyto-philic antibody (IgE) generated binds to mast cells Antigen binds to cell bound antibody crosslinks receptors, causing release of mediators Anaphylactic response to bee sting immediate response in allergic asthma... 

Quenching can actually be used to create analytical methods. An example is that of fluorescent antibody assay. Here, a fluor is bound to an antibody but the binding constant is less than that for the antibody-antigen pair for which the antibody was generated. Binding quenches the fluorescence of the fluor. When a solution containing the native antigen is mixed with the fluor complex, the fluor is released and can fluoresce normally. There are numerous examples of this approach. 

F(ab )2 fragments of antibodies are generated by pepsin digestion of whole IgG antibodies to remove a portion of the Fc region (see Sect. 1.1). Elimination of nonspecific binding of the Fc region of antibodies to Fc receptors on cell surfaces in... 

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

Fig. 25 The monoclonal antibody 26D9, generated to the jV-oxide hapten [67], catalysed the 6-exo-tet ring closure of [65] regioselectively to yield the disfavoured tetrahydropyran product [68]. This is a formal violation of Baldwin s rules, which predicts a 5-exo-tet spontaneous process to generate tetrahydrofuran derivative [66].

Fig. 34 (above) Antibody 38E1, generated from the a-hydroxyphosphonate hapten [99], catalysed the hydrolysis of p-nitrophenyl phosphate [100]. 

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... 

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