The Monoclonal Antibodiy Core Facility supports Memorial Sl0an-Kettering Cancer Center researchers by making it easier for them to generate and utilize monoclonal antibodies (MAbs) in their experiments. This is accomplished by being an informational resource as well as by providing various services.
Primary Services
The facility works with researchers to generate new custom MAbs by offering:
- Consultation on designing immunogens and relevant screening assays.
- Immunization of mice, rats or hamsters.
- Fusing B cells to myelomas to create immortal, MAb secreting hybridomas.
- Maintenance of the newly immortalized hybridomas during the screening process.
- Subcloning hybridoma populations that recognize the antigen of interest to create stable cell lines.
Memorial Sloan-Kettering researchers may search online in a growing bank of over 100 hybridomas the facility keeps in a general stock. MAbs are produced in vitro by cultivating the hybridoma at high density in CELLine Flasks (Integra Biosciences).
- Bioreactor Supernatants
- MAb concentration range is 0.3 to 4.5 mg/ml
- Production scale is 5 mgs to 1000 mgs
- Contain very low levels of bovine Ig
- Very low endotoxin levels (< 1 EU/ml)
- Stored sterile and azide-free
- Purified Antibody
- MAb concentration is always > 1 mg/ml
- MAb purity is usually > 95 percent
- Stored in PBS, sterile and azide-free
- Antibodies Conjugated to
- Alexa fluorescent dyes (488, 594, 633, 700, etc.)
- FITC, PE, APC
- Biotin
- HRP
- FAb and F(Ab')2 fragments
- Purified upon request
- Stored sterile and azide-free
The facility screens research samples weekly for the presence of mycoplasma / ureoplasma contamination.
Using a bioreactor system the facility produces secreted recombinant proteins from transfected CHO, 293 or myeloma cell lines, allowing production of proteins at higher concentrations than is normally obtainable using standard tissue culture techniques.
