At this stage

At this stage, when few weeks have passed by sample of blood are collected from the immunized mice and the antibody quantity is recorded. Loed and Quimbly quoted in 1999 that “several humane techniques have been developed for collection of small volumes of blood from mice.” Multiple methods can be used to identify the antibody titer and among these methods the ELISA and flow cytometry are the most popular. It is necessary for the titer to be in maximum amount in order for the fusion to be done. A boosting dose may be required in the case of insufficient titer amount. After the boosting dose blood was collected respectively until the enough repercussion is obtained. The boosting dose is also given even if the titer prevails in adequate amount. In this case the animals are treated solely by the antigen but this time no adjuvant will be used. This is done by either through intraperitonial route or IV route and must be performed 3 days prior to fusion but a couple of week followed by the initial antigen introduction. Then lastly the spleens from the immunized mice were collected in order to manufacture hybridoma cells in vitro.