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J Simková

Publications and source records attributed to J Simková.

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Attempt to isolate antierythrocyte antibodies by the method of affinity chromatography. II. IgG - incomplete antibodies.

It ensues from the results of the present work that erythrocyte stromata can serve as a suitable insoluble immunosorbent of both complete and incomplete antibodies. When applying optimum conditions, such as laboratory incubation temperature and suitable buffers, it is possible, by means of a chromatographic column with 0.5 ml erythrocyte stromata suspended in 40 g of glass beads (ballotini), to obtain fractions devoid of incomplete antibodies. Incomplete antibodies could be absorbed completely, either directly by a 24-hour incubation of the respective serum on the column or by passaging the serum through the column. In the presence of 0.2 M NaCl, absorption of incomplete antibodies was prevented; only complete antibodies were demonstrated in the fractions of the eluates. The elution yield in incomplete antibodies was low; in some cases, it could not be demonstrated at all. It can be explained partly by non-specific absorption, partly by the low initial antibody titre. It is therefore rather difficult to separate the individual incomplete antibodies from each other in this method. The column with incorporated erythrocyte stromata can be recommended for incomplete antibodies only because of its absorption capacity (to remove incomplete antibodies), but not for obtained elution yield.

Antibodies↗

Attempt to isolate antierythrocyte antibodies by the method of affinity chromatography. I. IgM--complete antibodies.

It follows from the results of the present study that erythrocyte stromata can serve as a suitable insoluble immunosorbent of complete antibodies. When using optimum conditions, such as laboratory incubation timperature, suitable buffers for both washing the column prior to absorption and for elution, it is possible to obtain, by means of the chromatographic column with 0.5 ml of erythrocyte stromata suspended in 40 g of silica glass beads (ballotini), fractions devoid of complete antibodies and subsequently to obtain the latter in purified form by elution. In complete anti antibodies, the elution yield amounted to an average of 35% of the initial amount of antibodies. Although the elution yield is not high and non-specific absorption has been demonstrated, the method can be recommended for the isolation of complete antibodies and thus for the production of monospecific sera because it is easy, rapid and inexpensive due to the regeneration ability of the mentioned columns and the relatively long duration of the immunoabsorption capacity of the incorporated erythrocyte stromata.

Absorption↗