Monoclonal Antibody Core
Under the new leadership of Edward Greenfield, the Monoclonal Antibody Core (MAC) has developed several new services to meet the specialized and growing demands of DF/HCC members and the greater Harvard medical community. In particular, the MAC has established technology and services to provide antibodies capable of immunohistochemistry, phospho-specific antibodies, flow cytometry, and are functional.
The Core provides expert consultation in antibody development to investigators and supports all aspects of antibody development, including generation of antigens such as peptides, recombinant protein, transfected cells, and cDNA; immunizations (subcutaneous, intraperitoneal, intramuscular, intrasplenic and intravenous) in standard 60 day or a rapid 21 day protocol; and antibody screening using ELISA, Western blot and flow cytometry formats. Multiple strains of mice, rats, and Armenian hamsters can be used to generate hybridomas and various knock-out and transgenic animals can be accommodated.
The MAC has established collaborations with 21st Century Biochemicals and Epitomics, Inc. to generate rabbit polyclonal and monoclonal antibodies. This includes a phospho-specific antibody package with double affinity purification of the sera to provide highly purified phospho-specific rabbit Ig. The rabbit polyclonal package includes an initial planning meeting, antigen design, five immunizations, serum titer and two production bleeds. The spleen can be cryopreserved for immediate or future monoclonal antibody generation.
The MAC offers in vitro monoclonal antibody production for any hybridoma generated or provided by the investigator. Hybridomas are grown up in either low Ig or serum-free, protein-free media in roller bottles or Integra flasks for yields of a few milligrams to hundreds of milligrams of purified antibody. Once purified, the MAC can also conjugate the antibody to different tags (biotin, HRP, AP, fluorochromes). Hybridomas can be provided by the investigator, purchased from the ATCC or generated by the MAC.
In addition, the MAC maintains a growing catalog of monoclonal antibodies against common antigens and biomarkers, like CD3 (OKT3; 145-2C11), CD4 (OKT4; GK1.5), CD8 (OKT8; TIB105), CD20 (B1), CD28 (EX5.3D10), CD80 (EW3.4B2; 16.10A1), CD86 (HF2.3D1; GL1), TcR (H57.597), c-myc (9E10), HA (12CA5) and GST to list a few. These are available in either conjugated or unconjugated form. The MAC can often provide these reagents at lower cost per milligram than they are obtainable commercially.
Every project begins with an initial planning meeting, in which the needs of the investigator are determined. Edward Greenfield, PhD is the Technical Director of the MAC, providing over 25 years of academic and industry experience in generating monoclonal antibody hybridomas. James DeCaprio, MD, serves as the scientific leader of the facility with 20 years experience generating monoclonal antibodies. Christina DeOssie is the Project Manager for the MAC. Together, the MAC has considerable expertise in antibody generation, antibody production/purification, recombinant protein technology, immunological assay development and molecular immunology. The Core is located at a Dana-Farberís North Campus facility, approximately 0.6 miles from the Longwood campus. Shuttle service is provided between the DFCI Main campus and the North Campus.