Hybridoma Re-fusion Introduction

Although hybridoma cell lines possess the characteristics of unlimited growth of myeloma cells, hybridoma cells that have been prepared for a longer period of time may still suffer from unstable growth and chromosome loss from one parent. In such cases, hybridoma re-fusion may be an alternative solution. The hybridoma re-fusion can add the corresponding parental chromosomal complement and thus achieve hybridoma rescue. Based on our well-established hybridoma platform, Creative Biolabs aims to provide you with the highest quality hybridoma re-fusion services to save your hybridoma and help your research.

Why is Hybridoma Re-fusion Technology Needed?

Hybridoma cells are obtained by fusing B lymphocytes with myeloma cells using a suitable screening method. Therefore, hybridoma cells theoretically possess both antibody-secreting function and immortality. However, the activity of hybridoma cells can be affected by the improper selection of methods, improper operation during the preparation of hybridoma, or even improper storage after the end of hybridoma preparation. In addition, hybridoma cells that have been prepared for a longer period of time can also suffer from instability and cell line phenotypic drift. These problems can lead to reduced activity of hybridoma cells and reduced efficiency of antibody production or even failure to produce the target antibody. Under these circumstances, hybridoma refusion can be a potential solution to rescue hybridoma cells and restore their activity.

How to Perform Hybridoma Re-fusion?

Hybridoma re-fusion involves fusing hybridoma cells with myeloma cell lines to obtain a new monoclonal cell line. The most suitable cell line is then selected among the new monoclonal cell lines for culture and antibody production. Specifically, this process includes several steps:

The first step is to select the appropriate myeloma cells. It's generally recommended to select two different myeloma cell lines for the re-fusion experiment according to the specific experimental situation to save time and ensure the success rate of re-fusion. The appropriate cell fusion method should be selected according to the specific experimental needs to perform the re-fusion of hybridoma cells and myeloma cells. Second, the fused cells are cultured and screened to screen out hybridoma-myeloma cell fusion cells. Then, the screened fusion cells are positively screened to identify positive cell lines that can express the target antibody. Finally, the positive cell lines are subcloned and the finalized positive cell lines are cryopreserved at the same time.

The most important step in the process is the selection of myeloma cell lines. There are three types of myeloma cell lines that can be used for hybridoma cell preparation, namely, mouse, rat, and human myeloma cell lines. Mouse myeloma cell lines include mouse P3-NS1/1-Ag4-1, mouse P3-X63-Ag8, mouse P3-X63-Ag8.653, mouse P3-X63-Ag8-U1, mouse SP2/O-Ag14, mouse 45.6TG1.7, mouse 45.6TG1.7.5, and mouse PuBul-Ou. Rat myeloma cell lines include rat Y3-Ag1.2.3. However, human myeloma cell lines currently may present difficulties such as low fusion rates, different growth rates in different media, slow cell growth, weak antibody secretion and low potency, and difficulty in cloning. Therefore, further research is required to improve human myeloma cell line usage.

Creative Biolabs has extensive experience in hybridoma re-fusion and can provide support and guidance to ensure the success of your project.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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