Simple Immunization Protocol for High Frequency Production of Soluble Antigen-Specific Hybridomas

Abstract

We report the immunization protocol used to produce high frequency of specific hybridomas secreting defined antibodies against soluble proteins. The two immunization schemes used consist in 1300 and 115 μg of protein distributed over a period of two weeks. The specific efficiency (SE) of positive clones recovered in eleven different fusion experiments ranged between 0.52 and 0.78. These values are referred to hybridomas secreting monoclonal antibodies (MAbs) against soluble proteins such as human chorionic somatomaminotropin (hCS), human growth hormone (hGH), and human prostatic acid phosphatase (hPAP). Similar high SE were also recovered by immunizing mice with nonsoluble antigens such as intermediate filaments of cytoskeleton (IF).