Immunization intrasplenic

Intrasplenic immunization is used for the production of hybridomas in situations where only very small quantities of the antigen are available. Typically, it lends itself extremely well to producing antibodies to proteins that have been purified by electrophoresis and subsequent blotting onto nitrocellulose. Antibodies produced by this route are always immunoglobulin class M as only one immunization is used. This method is covered by Animal Procedures legislation in most countries as it is an invasive surgical procedure and welfare issues must be addressed. 

Use the mice as donors for cell fusion 7 d after the intrasplenic immunization. 

In addition, improvements in immunization protocols such as suppression of dominant responses, in vitro or intrasplenic immunization, and antigen-targeting have also been developed. Suppression of dominant immune responses is used to permit the expression of antibody-producing cells with specificity for poor immunogens and is achieved using the selective ability of cyclophosphamide to dampen the immune response to a particular antigen, followed by subsequent immunization with a similar antigen. 

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