Bioassay for cell conditioned media
Inventors
Hnasko, Robert M. • Stanker, Larry H. • McGarvey, Jeffery A. • Lin, Alice V.
Assignees
US Department of Agriculture USDA
Publication Number
US-9868940-B2
Publication Date
2018-01-16
Expiration Date
2035-09-15
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Herein is described a novel hybridoma cell line (RMH359) and its use as a cellular bioassay to determine the efficacy of cell conditioned media (CCM) as a supplement used to support cell survival and promote growth in culture. This bioassay is the first to provide a measure of CCM bioactivity in support of hybridoma cells used in the generation and production of monoclonal antibodies.
Core Innovation
The invention describes a novel hybridoma cell line, RMH359, deposited as ATCC accession number PTA-122116, which requires macrophage conditioned media (MCM) for survival and growth in culture. RMH359 cells demonstrate MCM-dependent growth, allowing the use of this hybridoma line as a bioassay to assess the bioactivity of MCM, an undefined supplement used in hybridoma cell culture to support cell survival and promote growth.
The problem being addressed is the lack of standardization and guidance for the production and validation of MCM used as a supplement in hybridoma cultures. MCM is critical for supporting hybridoma survival and growth, particularly following cell fusion during monoclonal antibody production, yet its variable and undefined nature leads to inconsistent hybridoma viability and growth. This variability hinders optimization, quality control, and reliable monoclonal antibody generation.
The invention presents methods utilizing the RMH359 hybridoma cell line as a cellular bioassay to evaluate, standardize, and optimize conditions for MCM production. This includes determining optimal macrophage culture densities and times for generating MCM with maximal bioactivity, defining assay parameters such as MCM concentration ranges for supplementation, and validating MCM batches prior to culture use. The assay allows for the quantification of MCM bioactivity, facilitating improvements in hybridoma culture reliability and monoclonal antibody production quality.
Claims Coverage
The patent includes two independent claims encompassing the composition and a bioassay method related to the RMH359 hybridoma cell line and macrophage conditioned media.
Composition comprising conditioned media dependent hybridoma cell line and cell conditioned media
A composition including a hybridoma cell line deposited under accession number ATCC PTA-122116 that is dependent on conditioned media, specifically macrophage conditioned media, and the conditioned media itself.
Bioassay method for testing macrophage conditioned media bioactivity
A method involving: producing macrophage conditioned media; culturing the hybridoma cell line deposited under accession number ATCC PTA-122116; seeding these cultured hybridoma cells with the produced macrophage conditioned media; and measuring cell growth and survival to determine viability, thereby testing the macrophage conditioned media's ability to support hybridoma cell growth and viability.
The independent claims cover a novel hybridoma cell line composition requiring macrophage conditioned media for viability and a method of using this cell line as a bioassay to evaluate the bioactivity of macrophage conditioned media for supporting hybridoma cell growth.
Stated Advantages
Provides a standardized cellular bioassay for measuring the bioactivity of macrophage conditioned media used in hybridoma culture supplements.
Enables optimization and validation of macrophage conditioned media production to ensure consistent and maximal support of hybridoma cell survival and growth.
Facilitates batch-to-batch normalization of macrophage conditioned media, improving reproducibility and quality control in monoclonal antibody production.
Documented Applications
Use of the RMH359 hybridoma cell line as a cellular bioassay to evaluate, validate, and standardize macrophage conditioned media bioactivity as a supplement in hybridoma cell culture.
Application of the RMH359 bioassay to define optimal macrophage cell culture conditions for producing macrophage conditioned media with maximum efficacy in supporting hybridoma cells.
Use in the generation, selection, stabilization, and expansion of hybridoma cell lines producing monoclonal antibodies, particularly during early cloning stages requiring macrophage conditioned media supplementation.
Interested in licensing this patent?