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Hybridoma Re-Cloning Service

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Overview of Hybridoma Re-cloning

Hybridoma re-cloning is a popular hybridoma rescue measure, which aims to screen for highly active hybridoma populations. The best candidates should be stable manufacturers that allow them to demonstrate specific attributes based on product characteristics. Currently, limiting dilution, semi-solid medium, and automated system methods are the most representative strategies for hybridoma re-cloning. As a leader in the field of hybridoma research, Creative Biolabs has established a complete hybridoma screening platform to provide suitable hybridoma re-cloning pathways.

Why Implement Re-cloning?

It is well known that the antibodies produced by stable hybridoma cell lines will gradually decrease. Generally speaking, re-cloning hybridoma cell lines and selecting appropriate clones can effectively improve monoclonal antibody production. Simply put, hybridoma recloning is a powerful tool for retaining optimal clones.

What Can We Do?

Limiting dilution has long been considered a simple and easy-to-perform method for preparing positive hybridoma cells. In general, cells are diluted to an appropriate number per well, which contributes to increasing the proportion of wells containing a single clone. Several subsequent cloning procedures were performed until >90% of the wells contained the desired single clone. This strategy allows visual identification of cells. Smaller colonies represent slow-growing colonies while larger colonies represent fast-growing colonies.

Fig.1 Hybridoma cell screening. (Ao, Hang, et al., 2022)Fig.1 Assay of PCSK9-Ab secreted from hybridoma cells.1

As stable cell line technology matures, semi-solid media are emerging as an alternative to limiting dilution. The principle of semi-solid culture is similar to the process of colony screening. Hybridoma cells are first cultured and incubated on a specific plate for some time. After the desired colonies appear, the colonies are isolated for further screening. This strategy allows for cell fixation, which is extremely advantageous for tracking the growth of individual cells into clones.

In recent years, hybridoma screening and re-cloning technology based on automated systems has attracted increasing attention. This approach combines software and imaging systems that allow researchers to efficiently isolate and culture cells of interest. This technology optimizes the steps and time for re-cloning of hybridomas, which provides favorable conditions for efficiently and quickly selecting hybridomas and establishing stable cell lines.

Published Data

Hybridoma cell screening.Fig.2 Screening of specific hybridoma cells.1

Ao, Hang, et al. conducted tests on 32 individual hybridoma cell samples, each housed in separate wells, utilizing both chemiluminescence and ELISA techniques. Specifically, these methods were employed to screen for specific hybridoma cells based on their chemiluminescent properties. The supernatant of hybridoma cell samples, cultured for one day, was analyzed using the chemiluminescence method to detect the content of PCSK9-Ab. Additionally, the ELISA method was applied to measure the PCSK9-Ab content in the supernatant of hybridoma cell samples after six days of culture, along with assessing the cellular status at this time point. The results revealed significant disparities among the hybridoma cells in terms of their secretion and proliferation capabilities.

Why Choose Us?


Obtaining ideal hybridoma cells is a vital step in producing monoclonal antibodies. At Creative Biolabs, our professional technical team is dedicated to helping you re-clone and obtain high-yield cell lines to avoid losing valuable clones. If you are interested in more details of re-cloning services, please contact us in time.

Reference
  1. Ao, Hang, et al. "Chemiluminescent screening of specific hybridoma cells via a proximity-rolling circle activated enzymatic switch." Communications Biology 5.1 (2022): 308.
    Distributed under Open Access license CC BY 4.0, without modification.

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

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