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DFCI Monoclonal Antibody Core

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

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

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.

Affiliations:

People:

Resources:

Services

  • Antigen/recombinant protein development/cloning ( Material processing service )

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

  • Monoclonal Antibody Generation ( Material processing service )

    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 ( Material processing service )

    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 ( Material processing service )

    - 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 ( Material production service )

    Immunization
    - Rabbits (New Zealand White)

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


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Last updated: 2015-11-19T12:47:53.114-06:00

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The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016