METHOD OF PURE SPECIFIC ANTIBODIES PRODUCTION FOR PROTEINS CONCENTRATION'S QUICK QUANTITATIVE DETERMINATION

摘要

The production method for obtaining selective aggregations of antibody molecules of high purity and specificity, universally usable for a whole range of antibodies against normal proteins as well as pathological, depending on the type of the input material. A human serum, normal or pathological, or 10 percent water solution of lyophilized proteins or 10 percent water solution of lyophilized uric proteins or different substances, are treated by 1/100 to 1/10 of the volume of 25 percent glutaraldehyde water solution at 18 to 28 degrees C for the duration between 1 and 3 hours within the pH range of 8.0 to 9.0, from 1/20 to 1/5 of the volume of 10 percent water solution of potassium aluminium sulphate is added to the solution, and resulting floccules are to be isolated and washed by tap water. Afterwards, the floccules are submerged into three to ten-fold volume of a specific hyperimmune serum. After 1 to 3 days of observing the descent of the antibodies in the liquid the floccules are to be washed with tap water and eluated in 0.2M water solution of acetic acid with 2% of saccharose. Concentrated ammonia water is used to prepare the eluant for a neutral reaction, and the antibodies coagulate by dissolving 350 g of solid ammonium sulphate. After 16 hours of stabilization at 5 degrees C, the pure supernatant decants and the coagulated antibody dissolves in 0.15M of the water solution of sodium chloride, it dialysates and lyophilises. The obtained product is suitable wherever high sensitivity and speed is required for determination of normal and pathologic proteins, e.g. immunofluorescent, immunoadherent, nephelometric, and conjugation methods. As far as immunoprecipitation methods are concerned, it eliminates the elution of proteins from agar gel because it does not distort ballast protein background during the reactions.