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DFCI Monoclonal Antibody Core

Director: Greenfield, Edward A., Ph.D.

Location: 21-27 Burlington Ave., 4th floor, Boston, MA 02215

Core Summary:

The mission of the DFCI Monoclonal Antibody Core (MAC) is to provide immunological reagents (antibodies) to support research efforts of area investigators. We generate and produce novel monoclonal and polyclonal antibodies, recombinant proteins and transfected cells lines for use in basic research, drug discovery and clinical application, including diagnosis, surrogate markers for disease status, and response to therapy or drug toxicity.

Given the diverse research needs of the investigators at DFCI and the surrounding Harvard community, The MAC strives to support a wide range of antibody requests. A key component of the MAC is the ability to explore and develop new technologies that facilitate antibody generation. The MAC is set-up to provide all basic functions of antibody generation, screening, purification (milligram to gram amounts) and storing antibodies.

Institutions:
  • Dana-Farber Cancer Institute
Personnel/Contact Information:

Director: Greenfield, Edward A., Ph.D.
Phone: (617) 632-2209
Email

Member: Lama, Sujan, M.S.
Phone: (617) 632-7143
Email

Member: Bencsics, Craig, Ph.D.
Phone: (617) 582-8035
Email

Member: Bhanot, Haymanti, Ph.D.
Phone: (617) 632-9152
Email

Services:
  • Antigen/recombinant protein development/cloning

    Peptide synthesis
    Molecular cloning
    Recombinant protein development:
    - Mammalian expression
    - Baculovirus expression
    - Bacterial expression

  • Monoclonal Antibody Generation

    Immunization
    Animals (different species):
    - Mouse (different strains available)
    - Rat (different strains available)
    - Hamster (Armenian)

    Immunization schedules:
    - Standard (52-day; every other week)
    - RIMMS (21-day; every 2-3 days)
    - Custom

    Immunization types:
    - Standard immunization (SQ/IP + adjuvant)
    - cDNA immunization
    - Whole cell immunization
    - Subtractive immunization

    Hybridoma Fusion
    Fusion method:
    - PEG-assisted
    - Electrofusion

    Screening method:
    - ELISA
    - Western blot
    - Flow cytometry (FACS)

    Subcloning:
    - Limiting dilution plates

  • Monoclonal Antibody Production/Purification

    Production methods:
    - Standard (Roller bottle or T flask)
    - Integra flask
    - Fibercell hollow fiber

    Media:
    - Low Ig FCS medium
    - Serum-free medium

    Purification methods:
    - Standard (Protein A/Protein G)
    - Gel filtration (for IgM)
    - Low endotoxin

  • Other services

    - Isotyping
    - Mycoplasma testing (HEK Blue reporter assay)
    - Endotoxin testing (LAL)
    - Cryopreservation/Master cell banking
    - Productivity assay (Dot-blot)
    - Immunoassay development
    - Biacore affinity measurement
    - CDR sequencing

  • Polyclonal Antibody Generation

    Immunization
    - Rabbits (New Zealand White)

    Protocols
    - Standard (84-day; Pre-bleed + 4 production bleeds)
    - Phospho-specific (double affinity purification of sera)


Departmental Web Link(s):

Last updated: 2015-09-01T12:50:40.588-05:00