[Components of human complement system. Testing and isolation of C4]. / Komponenty sistemy komplementa cheloveka. Testirovanie i vydelenie C4.

A modified reagent for testing the hemolytic activity of human complement component C4 has been obtained. Reagent R4 was obtained by treatment of human blood serum pools with 0.075 M solution of hydrazine hydrate. This reagent was found to be rich in the serum fraction obtained by chromatography on DEAE-cellulose DE-52 and containing an active complement C3 component. To test the sensitivity and specificity of the reagent, component C4 was subjected to purification. This procedure resulted in a hemolytically active, electrophoretically and immunoelectrophoretically homogeneous component C4. DEAE-cellulose DE-52, DEAE-Sephacel, Ultragel AcA-34 and, again, DEAE-Sephacel were used consecutively as purification agents. The activity yield of component C4 with regard to the initial serum level was 20%.

[Components of human complement system. Testing and isolation of C3 and C5]. / Komponenty sistemy komplementa cheloveka. Testirovanie i vydelenie C3 i C5.

Modified reagents for testing the hemolytic activity of human complement components, C3 and C5, have been obtained. These reagents were obtained by treatment of human blood serum pools with a saturated solution of KBr (reagent R3) or 2 M KSCN and denaturated yeasts (reagent R5). These reagents were found to be rich in the serum factor obtained through the use of DEAE-cellulose DE-52 and containing the active component of the complement (C4). To test the sensitivity and specificity of the above reagents, components C3 and C5 were purified. After this procedure these components emerged as hemolytically active, electrophoretically and immunophoretically homogeneous components, C3 and C5. DEAE-cellulose DE-52, DEAE-Sephacel, Hydroxylapatite and Ultra-gel AcA-34 were used consecutively as purification agents. The activity yields of components C3 and C5 with regard to the initial serum levels were 31% and 18%, respectively.

Peculiar features of application of pH gradients formed in borate buffer with a polyhydroxy compound for separation of proteins in a free-flow electrophoretic apparatus.

The use of pH gradients in borate buffer with a polyhydroxy compound for separation of proteins in a free-flow electrophoretic apparatus is outlined. The composition of the pH gradients, the free-flow apparatus design and its operation, and the main causes of failures in the protein separation are considered.

Free-flow electrophoretic apparatus for separation and concentration of proteins.

Protein separation in the free-flow electrophoretic apparatus with 48 channels at both the inlet and outlet using "artificial" pH gradients was investigated. The separation was carried out in borate-mannitol pH gradients and in pH gradients created by the concentration gradient of boric acid in the solutions of borax and mannitol. The separation pattern depended on the ratio of the rate of sample injection to the flow-rate of solutions in the separation chamber and did not change when the protein concentration in the sample was changed. The protein separation in the pH gradient was better than in case of conventional free-flow electrophoresis. The free-flow electrophoretic apparatus with multi-channel inlet of the separation chamber is suitable for concentration of biological materials, e.g. proteins and bacterial cells.

[Evaluation of the mutagenic activity of chemical substances in an in vitro human peripheral blood lymphocyte culture after their activation in the body of mammals]. / Otsenka mutagennoi aktivnosti khimicheskikh veschestv v kul'ture limfotsitov perifericheskoi krovi cheloveka in vitro posle ikh aktivatsii v organizme mlekopitaiushchikh.

To evaluate mutagenic activity of human metabolites a method is suggested for treating the culture of human lymphocytes with whole mammalian blood after their exposure to a chemical. This method was used to study the cytogenic activity of cyclophosphamide metabolites available in blood of CBA female mice after intraperitoneal injection of different doses of the chemical.