Monoclonal Antibody Core Facility (DFCI)
9.5/10 Overall Rating
75% Positive
9 Orders Completed
Boston, Massachusetts, US
The Monoclonal Antibody Core at Dana-Farber Cancer Institute produces novel monoclonal antibodies for use in basic research, drug discovery, and clinical applications including diagnosis, surrogate markers for disease status, response to therapy, and drug toxicity. Services include cloning and scale-up of immunization antigens to purification of the resulting antibodies. Researchers can choose to have all phases of antibody production conducted by the core or perform particular steps in their own lab.
Services
Available animal models and cell lines:
We use standard strains of mice, rats and rabbits. Investigators can provide us with knock-out and transgenic animals when they are available. We have a variety of hybridoma fusion partner lines available (SP 2/0; P3X, NS1, YB 2/0, YB 1,2,3, 240E-1).
Equipment:
Biosafety cabinets, CO2 incubators, Molecular Devices Spectromax plate reader, Titertek plate washer, AKTA FPLC systems, access to Biacore and flow cytometers.
Lab members:
Edward Greenfield, PhD (Director); Craig Bencsics (Senior Scientist), Mohan Brahmandam (Scientist) and Yuhua Nong (Scientist).
Most recent publication:
Antbodies: A Laboratory Manual (edited by Edward Greenfield) published by Cold Spring Harbor Press 2013.
The Core is experienced in post-hybridoma antibody development and validation procedures that include isotyping, determination of antibody specificity, determination of CDR sequences of variable H and L chains and determination of antibody affinity by Biacore. Our core offers highly competitive prices for several antibody characterization services and significant discounts for multiple antibodies characterized and analyzed in parallel.
We offer extremely fast isotype testing service for hybridoma culture supernatants and purified antibodies. We also perform custom antibody specificity determination assays such as ELISA (both Indirect and Capture), western blot and FACS. The MAC also offers antibody validation services for purified antibodies, hybridoma culture supernatants and serum samples from immunized animals as well as custom biomarker validation and assay development services. We make every effort to validate antibodies by testing various conditions for biomarker studies.
Consultation on design of antigens and screening strategies
Additional costs for antibodies, ELISA kits, lysates.
Affinity chromatography (protein A and protein G)
We have an AKTA Express FPLC system.
Sample requirements depends on the type of antibody they need purified. We do both affinity (protein A and protein G) and size exclusion chromatography. We also can produce and purify recombinant proteins with Gst, His and Fc tags.
Turnaround time will depend on the volume or amount of material they are looking for. On average, a 100 ml scale mAb production takes about 3 weeks and 1 liter scale takes about 5 weeks.
We can accommodate projects up to a gram of low endotoxin antibody.
Cost is dependent on production scale of the hybridoma or recombinant cell line.
Animal immunizations: standard, RIMMS, subtractive protocols
Process Development describes the conversation we have with our clients to customize the steps that will be used in their specific project. This includes antigen design and production, animal species/strain selection and immunization protocol determination, assay selection and/or design, antibody characterization, hybridoma method selection and screening design, or something required that is unique.
Mammalian, baculovirus and bacterial systems
We have an AKTA Express FPLC system.
Sample requirements depends on the type of antibody they need purified. We do both affinity (protein A and protein G) and size exclusion chromatography. We also can produce and purify recombinant proteins with Gst, His and Fc tags.
Turnaround time will depend on the volume or amount of material they are looking for. On average, a 100 ml scale mAb production takes about 3 weeks and 1 liter scale takes about 5 weeks.
We can accommodate projects up to a gram of low endotoxin antibody.
Cost is dependent on production scale of the hybridoma or recombinant cell line.
Hybridoma fusion: PEG-assisted and electrofusion
"we were disappointed in the work product of the MAb Core at DFCI. The Western Blot they generated did not isolate for the right band of interest. They blamed the lysate but that has nothing to do with searching and isolating the appropriate band for the target. Not sure what happened but I wouldn't be inclined to use them again."
"Excellent. Provided high quality western blot service."
Monoclonal Antibody Core Facility (DFCI) has not received any endorsements.