The estimation of the amount of diphtheria toxin and its activity by various tests in the dynamic cultivation of Corynebacterium diphtheriae PW8]

Direct correlation between the results of tests for the biological activity of diphtheria toxin, carried out in vivo guinea pigs and in vitro in the microcytotoxicity test in CHO cell culture, has been established, which makes it possible to use the latter as one of the methods for the rapid, reproducible and economic evaluation of diphtheria toxin. The qualitative and quantitative evaluation of diphtheria toxin in the enzyme immunoassay with the use of monoclonal antibodies and in the microcytotoxicity test demonstrates that these two tests, when used for controlling cultivation processes, have essential advantages over the flocculation test as regards their specificity and information content.     

ATP-insulin conjugates and their use for immunofactor analysis

A method resulting in ATP-insulin conjugates by covalent binding of ATP modified at C(6) amino group of the adenine residue with insulin was developed. The modified ATP was bound to insulin by means of metha-p-toluene sulfonate-N-cyclohezyl Nf [2-morpholinyl(4)ethyl]-carbodiimide. The ATP analogs and ATP-insulin conjugates possess the coenzyme activity in a reaction of luciferin oxidation by luciferase from the fireflies Luciola mingrelica. the catalytic properties of soluble and immobilize on CNBR-activated. Sepharose enzymes in reactions with native ATR, its modified derivatives and ATP--insulin conjugates were compared. The bioluminescence reaction involving ATP--insulin conjugate is inhibited by antibodies against insulin. This effect can form a basis for insulin detection in solution, which is based on competitive binding of free and antibody-labelled ATP--insulin conjugates.