Spleen cells preparation from immunized mouse

Catalytic antibodies were first prepared in the mid-1980s. In one example, a rabbit or mouse was immunized with phosphonate ester A, and pure anti-A antibodies were isolated from this animal s spleen cells. Some of the anti-A antibodies were found to catalyze the hydrolysis of the ester functionality of B at rates significantly faster than background. 

The volume of the tissue should not exceed 10% of the volume of TRI REAGENT . Alternatively, rather than using the whole spleen directly, splenocytes can first be prepared from spleen by using standard procedures described for preparation of hybridoma production (see Chapter 1) before total RNA extraction. A spleen from an immunized mouse contains approx. 5 X 10 to 2 X 10 lymphocytes. For total RNA extraction, substitute 10 lymphocytes for a spleen and simply add 10 ml of TRI REAGENT (1 ml of reagent for 10 cells) to the sterile PBS-washed cell pellet. Allow incubation at room temperature for 5 min for lysis, and proceed to Protocol 2. step 2. 

MAbs were first produced by mouse hybridoma cells prepared by fusing spleen cells from an immunized mouse with mouse myeloma cells. The hybridoma cells retain the capacity to make specific antibody while the myeloma cells impart the capacity of the cells to grow indefinitely in culture, continuously seaeting antibody. 

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