Inactivation conjugate

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Classify each of the routine vaccines as an inactivated, polysaccharide, conjugate, toxoid, or subunit vaccine. 

It has become apparent that the method of crosslinking can dramatically affect the activity of an immunotoxin in vivo. This is true not only with regard to possible direct blocking by the crosslinker of the enzymatic active site which is responsible for inactivation of ribosomes, but the chemistry of conjugation also is an important factor in proper binding and entry of the... 

Concentrate the thiolated toxin to lOmg/ml using centrifugal concentrators. Immediately use the modified protein in the conjugation reaction to prevent inactivation of 2-iminothiolane-modified molecules by recyclization (Chapter 1, Section 4.1). 

Intracellular thiolate ligands such as glutathione (GSH, the tripeptide y-L-Glu-L-Cys-Gly) are believed to inactivate cisplatin because the reactions with cisplatin tend to be irreversible (35). Elevated levels of GSH have been observed in cisplatin-resistant cells. Recently, it has been shown that an MRP gene, which encodes a human ATP-dependent glutathione S-conjugate export pump (GS-X pump), is expressed at higher levels in cisplatin-resistant (HL-60/R-CP) cells than in sensitive cells (36). The GS-X pump may contribute to the excretion of Pt-GS complexes from cells (37). 

Another example that can be included in the subunit vaccine class is the use of bacterial toxoids. Many bacteria produce toxins which play an important role in the development of the disease caused by a particular organism. Thus, vaccines against some agents, for example tetanus and diphtheria, consist of the toxin inactivated with formaldehyde conjugated to an adjuvant. Immunization protects from disease by stimulating antitoxin antibody which neutralizes the effects of the toxin. 

Ready-to-use blocking solutions containing normal inactivated sera or BSA for use with colloidal gold conjugates are available from Aurion (http //www. aurion.nl/). When labeling with protein A and protein G conjugates, this step can be omitted. 

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