Radiolabel chemical labeling

Radioactive tracers account for about 20% of the worldwide market for consumables and reagents for life science research. In 1994 the value was estimated at about 300 million. The principal fuU line manufacturers are Du Pont—NEN Research Products (Boston, Massachusetts) and Amersham International (Amersham, U.K.). These companies share roughly equaHy about 85% of the radiochemicals worldwide market. In addition to an extensive line of catalog products, these suppHers offer custom labeling and custom synthesis services. The rest of the market is shared by producers of a limited range of products or services, such as ICN Biomedicals (Costa Mesa, California) and American Radiolabeled Chemicals (St. Louis, Missouri). 

Armexin A1 emerged as a promising antigen for the radiolabeled antibody-based imaging and therapy of cancer [100]. In our laboratory, we use terminal perfusion protocols featuring active esters of biotin for the selective chemical labeling of accessible proteins in vascular structures. Biotinylated proteins are then purified from different organs (collected separately) and are submitted to a comparative proteomic analysis [106]. 

Muelder and Shadoff (3) prepared C-2,3,7,8-Cl4-DBpD (0.9 mCi/ mmole) by chlorination of C-2,7-dichlorodibenzo-p-dioxin made from potassium C-2,4-dichlorophenate. The preparation of tritium-labeled 2,3,7,8-Cl4-DBpD is justified because the radiolabeled intermediates are less expensive and more accessible and because a higher specific activity is potentially attainable. Here, we consider the optimal conditions for the reaction sequence designed to obtain products of high chemical and radiochemical purity shown at the top of p. 8. 

Typical validation for radiochemical and radiopharmaceutical purity. Quality control is very important to ensure the safety and efficacy of radiopharmaceuticals. One important quality parameter is the radiochemical purity of the radiolabeled product. This is defined as the fraction of the total radioactivity in the desired chemical form in the radiopharmaceutical [56]. Radiochemical impurities come from incomplete labeling, shift of equilibrium, radiolysis ((3 decay), temperature or pH change, exposure to light,... 

Expired air. For 14C-labeled chemicals, the tracer carbon may be incorporated in vivo into carbon dioxide, a possible metabolic product. Therefore, when the position of the radiolabel indicates the potential for biological instability, a pilot study to collect expired air and monitor its radioactivity content should be conducted prior to initiating a full-scale study. Expired air studies should also be performed in situations where the radiolabel has been postulated to be stable but analyses of urine and feces from the toxicokinetic study fail to yield complete recovery (mass balance) of the dose. 

Many of radioactive isotopes are very useful for the following biochemical processes (Table 6.1). The radioactive label is introduced into macromolecules, especially proteins, either during biosynthesis, e.g., during translation in the presence of S-methionine, or enzymatically, e.g., by use of P-labeled ATP during protein phosphorylation by protein kinases, or chemically by modification of amino acid side chains. Examples for reagents used in chemical radiolabeling of proteins are given in Table 6.2. 

To make these substrates suitable for biological assays, the introduction of functional groups that can be traced with the proper analytical techniques is essential. The use of radio-, fluorescent-, and biotin-labeled lipidated peptides has been reported. The synthesis of fluorescent substrates is chemically straightforward and allows for production of larger quantities than the enzymatic synthesis used for radiolabeled peptides and is thus preferred over the use of radioactive compounds. [1 21] Common fluorescent probes can be introduced by conjugation to a free functional group present in the peptide. The fluorescent moiety is... 

Chemical synthesis of labeled compounds suffers from some limitations and problems, though. One limitation concerns the amount and cost of the radioactive starting material. This factor necessitates devising synthetic routes to the desired compounds in which the radiolabel can be introduced near the end of the sequence of reactions, so as to secure as high an overall yield of labeled material as possible. At present, numerous labeled compounds are available commercially as starting materials for syntheses. Still, in planning a new synthetic route, it is necessary to consider its compatibility with the specific stalling material available. 

The chemical purity of these radiolabeled compounds is very high from most sources. If in question, the purity can be checked and restored by standard TLC and/or HPLC methods. 14C-labeled phytosterols are less available. 

The combination of radiolabeled sulfide and the bimane-HPLC method is particularly powerful because one of the main obstacles to the use of labeled sulfide is, that aside from radioactive decay, the compound is subject to rapid oxidation in the presence of air. The breakdown products of chemical sulfide oxidation are the same as those of biological oxidation. Previously it has been impossible to check routinely the purity of the purchased isotope and its subsequent purity during a series of experiments. It is our experience that newly purchased sodium sulfide sometimes contains up to 10% thiosulfate as well as traces of sulfite and sulfate (Figure 2), and that the sulfide once hydrated readily oxidizes if stored in a normal refrigerator. 

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