Aminoethyl Amino Research Articles

Effectiveness of Cytoprotective Agents on Sulfur Mustard Induced Toxicity: The In vitro Model

Sulphur mustard (bis (2-chloroethyl) sulfide, SM) is a powerful vesicating chemical warfare agent that causes profound injuries to the eyes, lungs and skin. Despite intensive research following the first use of SM in World War I, there is still no useful pretreatment or therapeutic antidote available. This agent remains a constant chemical threat. A potential approach to combating the debiliting effects of this agent is the use of compounds that can react with this material before it interacts with critical macromolecules. Glutathione (GSH), a tripeptide that exists in high concentrations in cells, reacts with SM and is involved in SM detoxification. Amifostine is a synthetic aminothiol, has been extensively used as a radioprotector. This prompted us to evaluate the protective efficacy of GSH, Amifostine and DRDE-07 (S-2(2- aminoethyl amino) ethyl phenyl sulfide) (synthesized in our lab) against SM toxicity in vitro in HeLa cell line. All these compounds are thiol group containing compounds. Pretreatment of HeLa cell with these cytoprotectants led to decrease in cytotoxicity after SM exposure. The protective efficacy of above compounds were evaluated against sulphur mustard using HeLa cells. The above compounds were added to the media 1 hr before the SM exposure and incubated for 24 hrs. cell viability by MTT assay and LDH leakage were measured as end point.

Immunoadsorbents for the isolation of high-affinity anti-hapten antibodies in high yield

Anti-hapten antibodies with affinities close to 10 8/M can be isolated in high yield by elution under mild conditions from immunoadsorbents prepared by coupling haptens directly to an aminoethylamino (AEA) derivative of beaded agarose. In experiments with rabbit anti-2,4-dinitrophenyl (DNP) antibody with an average association constant ( K 0) of 1.2 × 10 9/M, elution of antibody bound to a Sepharose-AEA-DNP column produced a yield of 57%. With rabbit antibody to the 5-dimethyl-aminonapthalene-1-sulfonyl (DANS) hapten with K 0 = 2.7 × 10 8/M and a Sips heterogeneity index a = 0.52, a 20% yield of the antibody found in the starting material by quantitative precipitin analysis was obtained by specific precipitation and dissolution of the precipitate with 0.18 M Na DANSulfonate; this antibody had K 0 = 6.6 × 10 8/M and a = 0.66. Purification of another aliquot of the same antibody preparation by immunoadsorption on DANS-AEA-Sepharose and elution with the same hapten concentration produced a yield of 75% of antibody with K 0 = 5.7 × 10 7/M and a = 0.29. In 3 production runs with the new method, apparent yields of 181, 101 and 94% (based on quantitative precipitin analysis of the starting materials) of anti-DANS antibody were obtained. In 2 of these runs, the immunoadsorbent was further eluted with 0.1 M acetic acid and then with 0.1 M acetic acid in 10% dioxane; additional yields of 9 and 4% were obtained with acetic acid and further yields of 15 and 11% with acetic acid/dioxane; the antibody eluted with acetic acid was damaged as shown by its failure to mediate passive immune lysis of DANS-coupled sheep erythrocytes, but the antibody eluted with acetic acid/ dioxane was recovered intact.