Purification of mouse myelin basic proteins by immunoaffinity chromatography.
The preparation of two immunosorbent columns specific for mouse myelin basic proteins (BPs) is described. The columns were prepared from either affinity purified mouse 14K-antibodies or bovine BP fragment (residues 1-115) antibodies. The column prepared from mouse 14K-antibodies bound 216-441 micrograms of mouse BPs within its working range at pH 4.5. The second column, prepared from antibodies specific for bovine fragment 1-115, bound 245-442 micrograms of mouse BPs under similar conditions. The ratios of 14K to 18.5 K proteins present in the immunopurified BPs reflected those in the samples originally applied onto the column. Therefore, both columns were equally effective for the small scale isolation of mouse BPs from whole brain. Because it is easier to prepare large quantities of bovine BP fragment 1-115 than the mouse 14K protein, it is less difficult to obtain antibodies against the bovine fragment. As a result, the use of immunosorbent columns containing immobilized bovine BP fragment 1-115-antibodies to purify mouse BPs represents a practical alternative to using immunosorbent columns prepared with mouse 14K-antibodies.
D000818:Animals; D001923:Brain Chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D007163:Immunosorbent Techniques; D051379:Mice; D004676:Myelin Basic Protein / Q000302:isolation & purification*
D004676:Myelin Basic Protein