Customer Case | How does InnoMedia® Hybridoma Basal Medium help achieve efficient cell culture?

Background Introduction

The production of murine monoclonal antibodies (mAbs) using hybridoma technology has long been a crucial platform for therapeutic development and diagnostic applications. However, with the advancement of CHO antibody production platforms, hybridoma technology is now more commonly used for rapid antibody generation in the early stages of research and development. This shift has posed new challenges for hybridoma basal media, which must demonstrate superior serum-free adaptability and be specifically optimized for batch culture.

Traditionally, the serum-free adaptation of cells cultured in serum-containing media often leads to a decline in cell performance and productivity. Our InnoMedia® Hybridoma Basal Medium addresses these challenges by offering excellent adaptability, enabling direct seeding and culture from serum-containing or other serum-free media with almost no need for adaptation. Furthermore, it supports the direct transition from adherent culture to suspension culture without requiring cell adaptation.

InnoMedia® Hybridoma Basal Medium is a chemically defined, low-protein, animal-component-free basal medium suitable for the suspension culture of hybridoma cells and myeloma cells. It supports hybridoma cells from the adaptation stage to high-yield antibody production in batch culture.

Key advantages include:

• Passage stability: Supports culture for more than 40 passages/3 months.
• Serum-free adaptability: Enables direct transition from traditional serum-containing medium to suspension culture without the need for serum-free adaptation.
• Specialization for batch culture mode: Compared with competing media, the nutrient content in the medium is enhanced, eliminating the need for additional feeding during the batch culture cycle.
• High cell viability and antibody yield: Improves cell growth and IgG production during the batch culture cycle.

Direct Hybridoma Monoclonal Antibody Production from Adherent Culture to Suspension Culture

This application note demonstrates the excellent performance of our hybridoma CD basal medium in monoclonal antibody production, as well as the direct transition from adherent culture to suspension culture, with a focus on its impact on cell growth, viability, and IgG productivity in a single batch culture.

Materials and Methods:

• Cell lines: Hybridoma cell lines A & B (Sp2/0-derived cell lines, producing monoclonal antibodies with a molecular weight of approximately 150 kDa).

• Initial culture conditions: Cells were cultured in DMEM + 10% FBS adherent culture, then directly transferred to suspension culture using CD hybridoma medium.

Culture Conditions:

• Basal media:

• InnoMedia® Hybridoma Basal Medium (CM-L101001) CD hybridoma media from competing companies #1, #2, and #3

• Culture system: 125 mL plastic disposable shake flasks (with a culture volume of 45 mL) were used for cultivation on an orbital shaker platform at 130 rpm.

• Cells were seeded at 1 × 10^6 viable cells/mL. All cultures were maintained in a humidified atmosphere with 8% CO₂. No feeding: InnoMedia® medium required no additional feeding during batch culture, while other media were supplemented as necessary according to the manufacturers' instructions.

In the comparative cell growth study, hybridoma cells grown in DMEM + 10% FBS were directly seeded into various hybridoma CD media to evaluate growth and monoclonal antibody production in direct suspension batch culture. Cell counts and viability were determined using trypan blue staining with the Altair™ automated cell counter (Countstar™). Antibodies were collected via single-step purification with Protein G, and the titer of purified IgG was measured using NanoDrop™ based on the calculated extinction coefficient.

Experimental Results:

In the case of Cell Line #1, the peak viable cell density (VCD) of our medium reached 6.7 × 10^6 viable cells/ml, which was approximately 200% higher than that of the best competitor tested; the IgG titer reached 170 mg/L, which was 170% higher than that of the best competitor tested.

   In the case of Cell Line #2, the peak viable cell density (VCD) of our medium reached 13.2 × 10^6 viable cells/ml, approximately 320% higher than that of the best competitor tested; the IgG titer reached 250 mg/L, 250% higher than that of the best competitor tested.