Single-Cell Cloning of Hybridoma Cells by Growth in Soft Agar

Author:

Greenfield Edward A.

Abstract

Single-cell cloning during hybridoma production ensures that cells that produce the antibody of interest are truly monoclonal and that the secretion of this antibody can be stably maintained. Cloning of hybridoma cells in semisolid medium is one of the most commonly used methods for producing single-cell clones. The technique is easy, but, because it is performed in two stages, it does take longer than other methods. Not all cells will grow in soft agar, and there may be a bias on the type of colony that appears. However, most of the commonly used myeloma fusion partners have relatively good cloning efficiencies in soft agar, and, consequently, so do most hybridomas. Even though every attempt is made to ensure that the cells are in a single-cell suspension before plating, there is no way to guarantee that the colonies do not arise from two cells that were stuck together. Therefore, single-cell cloning in soft agar should be repeated at least twice before the cells are considered clonal.

Publisher

Cold Spring Harbor Laboratory

Subject

General Biochemistry, Genetics and Molecular Biology

Cited by 4 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

1. Generating Monoclonal Antibodies;Cold Spring Harbor Protocols;2022-08

2. Growing Hybridomas;Cold Spring Harbor Protocols;2021-10

3. Ridding Hybridoma Cell Lines of Contaminating Microorganisms;Cold Spring Harbor Protocols;2021-07

4. Preparing Feeder Cell Cultures to Support Hybridoma Growth;Cold Spring Harbor Protocols;2019-11

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