Hybridoma Core

Services

Antigen Conjugations

Peptide conjugation to KLH by a glutaraldehyde protocol.

Cloning

Limiting dilution cloning of five chosen hybridoma cell lines. Collection of media samples for screening by the investigator. Minimal scale up and preparation of frozen stocks (5 vials) of clones are completed by HCF. Includes labor and supplies. This phase goes 4-8 weeks.Freezing Cells Cells from culture expansion associated with mAb production can be frozen down for storage in Liq N2. The freezing media is 90% FBS:10% DMSO and the cells are at a concentration of 2-4 x 106 cells/ml.

ELISA

We can perform antigen-capture or sandwich type ELISAs to measure antibody levels in tissue culture supernatant.

Freezing Cells

The Core will freeze back hybridomas in freezing media from actively growing cells.

Fusion of cells

Fusion of spleen cells from chosen mouse to myeloma cell line. Plating of fused cells and collection of media samples for screening by the investigator. Includes labor and supplies. This phase goes 4-6 weeks.

Integra Culture System

For production of mAb from cell lines This system is intended for production of 30-60mg of mAb per month in the smaller system and 100-200mg of mAb in the larger system. Ave. concentration is 1.5mg/ml. The production schedule ave. is 8 weeks.

Large-scale Antibody Production

The Hybridoma Core uses a static cell culture system the Integra Flask to produce high concentration (>.5mg/ml) monoclonal antibodies. This can be done in serum-free or using ultra-low IgG/IgM serum. Yields can be as high as 100mg/month/flask.

Large-scale antibody purification

We can purify up to 80mg of IgG from one sample using a protein G column.

Liquid Nitrogen Storage of cells

Storage for cloned cell lines. The stored cell lines must be mycoplasma free.

Mouse injections

We will perform intraperitoneal injections of antigen into mice to elicit an immune response for the production of monoclonal antiodies.

Polyclonal Antibody Production

This service will be offered on a limited basis depending on available housing. One rabbit per antigen. Includes animal purchase and board, 6 injections, 1 test bleed, 3 production collections, final bleed and labor for 120 days. Time course can be extended for additional fee based on animal board and labor charges. The goal is to provide a total of 100mls of serum, but this is not guaranteed. Allow 1-2 weeks for animals to arrive.

Anti-thymocyte globulin (ATG) in Rabbit

We reliably produce mouse anti-thymocyte globulin in rabbits using a proprietary method of mouse thymocyte antigen preparation.
(This serum is for research purposes only).

Purifications

We can purify monoclonal or polyclonal antibodies using Protein G or an epitope specific affinity column.

Western Blot

We will run the SDS PAGE gel, transfer and detect using investigator provided primary antibody plus ECL reagent.

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