||Improve Serum-Free Hybridoma Culture with Gibco® Media
|Product||CD Hybridoma Medium✝||CD Hybridoma AGT™✝✝||Hybridoma-SFM✝||PFHM-II|
|Media Type||Chemically-Defined Media||Chemically-Defined Media||Serum-Free Media
Low-protein 20 μg/ml
|Optimized For||Human, mouse, rat hybridomas,myelomas. NS0, NS-1, and other steroid-dependent cells when used with 250X Cholesterol Lipid Concentrate||Human, mouse, rat hybridomas, myelomas|
|Applications||Growth and MAb production. Can be used to express other proteins in engineered myeloma cell lines.|
✝Drug Master File available
✝✝Dry granular format of CD Hybridoma Medium
Note: Cell lines from different sources, and different clones of the same cell line, may have highly specific nutritional requirements and may therefore prefer one medium over another. More than one medium formulation (if available) should be evaluated to determine the best option.
Gibco® CD Hybridoma Medium is a chemically defined, protein-free medium optimized for the production of monoclonal antibodies and recombinant proteins in human, mouse, and rat hybridomas and myelomas. Because it is manufactured with animal origin–free materials, protein purification and downstream processing of cultures grown in CD Hybridoma Medium is streamlined compared to that required for cultures grown in less defined media. It is also less likely to contain adventitious agents that can compromise your cultures. CD Hybridoma Medium is formulated without L-glutamine, providing added stability in culture.
Suitable for the culture of recombinant myeloma lines as well as traditional hybridomas, CD Hybridoma Medium outperforms serum-free and serum-supplemented hybridoma media in terms of cell culture density and IgG production (Figure 1). When supplemented with 250X Cholesterol Lipid Concentrate, this medium is also well suited for lipid- or cholesterol-dependent cultures such as NS0-derived lines. Cells growing in traditional serum-supplemented medium can be adapted easily to this medium, typically in as few as 3 passages. Whether you’re growing in stationary flasks, agitated suspension systems, or even bioreactor-scale culture systems, CD Hybridoma Medium supports maximum cell densities and high protein yield.
CD Hybridoma Medium is available in a ready-to-use (1X) liquid format and in our easy-to-use Advanced Granulation Technology™ (AGT™) format. AGT™ media are complete and pH-adjusted; L-glutamine-dependent systems require only standard supplementation with L-glutamine (or GlutaMAX™-I Supplement). The granules dissolve instantly for faster media preparation compared to traditional powdered media. AGT™ media can help reduce total cycle costs, decreasing time required for raw material planning, procurement, and testing, as well as media preparation.
Growth of Hybridoma TP4 - 3.1
InG Production by Hybridoma TP4 - 3.1
|Figure 1. Gibco® CD Hybridoma Medium results in higher viable cell densities and antibody yield in Hybridoma TP4-3.1 cells than other media.
In comparative cell growth studies, Hybridoma TP4-3.1 (proprietary) cells growing in IMDM + 10% FBS were adapted to growth in other media through a minimum of 3 passages prior to evaluation of growth and monoclonal antibody production in batch culture.
CD Hybridoma Medium was supplemented with 8 mM L-glutamine prior to use. Other media were supplemented as necessary following manufacturers’ instructions. Agitated small-scale TP4-3.1 suspension cultures were grown in 125 mL plastic disposable shake flasks (35 mL culture volume) on an orbital shaker platform at 125–135 rpm. Cells were seeded at 105 viable cells/mL. All cultures were incubated in a humidified atmosphere of 8% CO2 in air. Total cell counts were determined using an electronic particle counter; viability of cells was estimated by trypan blue dye exclusion. IgG was measured by ELISA.
A–Gibco® CD Hybridoma Medium
D, E and F are media from other manufacturers.
PFHM-II is a protein-free medium optimized for the growth of human, mouse, and rat hybridomas and myelomas, and the production of monoclonal antibodies. It contains no polypeptides such as growth or attachment factors, which could interfere with downstream protein purification. PFHM-II performs as well as serum-supplemented media for monoclonal antibody production.
Hybridoma-SFM is a very low-protein medium (20 μg/mL) optimized for the growth of human, mouse, and rat hybridomas and myelomas and the production of monoclonal antibodies (mAb). It provides excellent growth and maintenance of hybridoma cells and offers significantly higher mAb yields and simpler downstream processing than do serum-supplemented media (Figure 2). A complete, ready-to-use medium, it is supplemented with trace elements, minerals, and a low amount (20 μg/mL) of defined protein (insulin and transferrin).
Unlike serum-supplemented media, Hybridoma-SFM is free of BSA, steroids, and endogenous bovine immunoglobulin, facilitating purification of specific mAbs. It is easy to use and supports a wide range of hybridoma cell lines in a variety of culture systems. Most hybridomas require only minor sequential adaptation or no adaptation to this medium.
Cumulative MAb Production and Number of Cells
|Figure 2. Gibco® Hybridoma-SFM supports higher yield of monoclonal antibodies from comparable cell quantities than D-MEM.
Cells were cultured in Hybridoma-SFM or D-MEM with FBS (10% until day 10, 7.5% until day 13, and 5% from day 13). Cell number and mAb production were monitored. The culture grown in Hybridoma-SFM produced significantly more mAbs even though the number of cells was similar to that of the culture grown in serum-supplemented medium.