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.
D000906:Antibodies / Q000302:isolation & purification*; D000917:Antibody Formation; D002846:Chromatography, Affinity; D003298:Coombs Test; D004912:Erythrocytes / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D013696:Temperature
D000906:Antibodies; D007074:Immunoglobulin G