Hybridoma

Overview

The Hybridoma Facility will produce monoclonal antibodies to antigens provided by investigators. For most investigators, the staff of the facility will carry out the whole process from immunization through the final cloning, freezing and production of mAb. In general Dr. Scharff and Ms. Buhl will meet with the Principle Investigator, or his or her representative, to discuss the scientific problem and make decisions about how to immunize animals, to screen for positive hybridomas once a fusion is done and on the types of antibodies that are required. In cases where the screen for positive hybridomas requires special expertise, someone from the PI’s lab participates in the initial screening. Subsequent cloning and freezing is done by the facility and key clones are stored as a backup for investigators. In the course of making the hybridomas, students and fellows can receive training on how to carry out the hybridoma technology. We also provide advice and letters of support for grant applications that involve making and using monoclonal antibodies.

Services

Decisions on optimum antigens and immunization protocols
Immunization of mice and bleeding and assaying mouse serum for titers and characteristics of antibodies
Selection of best mouse for fusion and the fusion of mouse myeloma and immune spleen cells
Repeated screening of ~20 96-well plates for positive hybridomas
Cloning and recloning to stabilize positive hybridomas
Freezing of positive hybridomas
Generation of mg/ml concentrations of monoclonal antibodies where required