How do you select the myeloma fusion partners for my specific hybridoma?

We maintain a panel of well-characterized, high-performance myeloma cell lines. The selection process considers the species of origin of your hybridoma (e.g., mouse, rat), any available data on its previous fusion partner if known, and the growth characteristics desired. Using at least two different myeloma lines in parallel increases the chances of finding a highly compatible and efficient partner.

My original hybridoma is in very poor condition (low viability, no detectable antibody). Is re-fusion still an option?

Even if your hybridoma is in poor condition, re-fusion may still be a viable option, provided some viable cells are remaining that harbor the genetic material for your antibody. The re-fusion process is designed to provide a new, healthier cellular environment. We recommend discussing the specifics of your cell line with our experts for a thorough evaluation.

What if multiple stable, high-producing clones are rescued from the re-fusion? Do I get all of them?

Yes, if our re-fusion and subcloning efforts yield multiple distinct clones that meet the criteria for stability and high antibody production, we will discuss these findings with you. Typically, we will provide you with an agreed-upon number of the best-performing clones (e.g., top 2-3) as part of the standard service, with options to receive additional characterized clones if desired.

How is the antibody production of the rescued clones confirmed and quantified?

Antibody production is initially confirmed through screening assays like ELISA or Western blot, using culture supernatants from emerging clones. For lead candidate clones, we can perform more quantitative assessments of antibody secretion rates (e.g., µg/ml/10^6 cells/day) to ensure they meet your productivity requirements. The specific assays can often be tailored using antigen or detection reagents you provide.

What are the final deliverables I will receive after completing the service?

Upon completion, you will receive cryopreserved vials of the successfully rescued and subcloned hybridoma cell line(s) – typically 2-5 vials per selected clone. You will also receive a comprehensive final report detailing all methodologies, screening data, subcloning results, and characterization of the rescued clones, including viability and antibody productivity assessments.

Hybridoma Re-Fusion Service

Overview Service Features FAQ

Creative Biolabs offers a specialized Hybridoma Re-Fusion Service designed to rescue your valuable yet underperforming hybridoma cell lines. With our extensive experience and cutting-edge facilities, we can restore antibody production and ensure the continuity of your critical projects, effectively safeguarding your scientific investment and progress.

Overview of Hybridoma Re-Fusion

Cell fusion, the process of merging two distinct cell types into a single hybrid cell, can be achieved through natural or artificial means, resulting in progeny that exhibit characteristics from both parental cells. This technology is fundamental to modern biotechnology, particularly in the generation of monoclonal antibodies (mAbs). The creation of mAbs necessitates the fusion of antibody-producing cells with continuously dividing cancer cells, generating hybridoma cells.

An electrofusion platform. (Wu, et al., 2022) Fig.1 Schematic of cell fusion platform.¹

Hybridoma technology remains a widely employed conventional method for mAb isolation. Its significant advantage lies in the continuous production of mAbs once stable hybridoma cell clones are established. This method involves fusing immortalized cells with antigen-specific B cells, thereby facilitating B cell culture and enabling persistent antibody secretion. However, prior cytogenetic studies have revealed a propensity for chromosome loss from one parental cell during continuous proliferation. This chromosomal instability can lead to compromised cell growth, reduced expression levels of the target protein, and a decline in product quality. Furthermore, suboptimal preparation methods, inaccurate operational procedures, or inappropriate storage conditions can adversely affect hybridoma viability and mAb production efficiency. Consequently, timely hybridoma re-fusion emerges as a promising method to address these issues.

Hybridoma re-fusion involves fusing existing hybridoma cells with myeloma cell lines to generate a new monoclonal cell line. The most suitable cell line among these newly formed hybridomas is then selected for subsequent mAb production. This technique has been recognized as an effective strategy to counteract chromosome loss by augmenting the corresponding parental chromosome complement, thereby aiding in the rescue of compromised hybridoma cells and the restoration of their activity.

Our Hybridoma Re-Fusion Service

At Creative Biolabs, we know myeloma fusion partner selection is key for successful hybridoma rescue. We re-fuse your hybridoma with at least two distinct, high-performance myeloma cell lines from our extensively characterized panel. This approach helps us establish new monoclonal cell lines, rigorously selected for restored antibody production, optimal growth, and long-term stability, ensuring you receive a revitalized, reliable resource for your research.

Workflow of hybridoma re-fusion. (Creative Biolabs Original)

01Strategic Myeloma Cell Line Selection

We initiate the process by carefully selecting appropriate myeloma fusion partners from our proprietary, well-characterized bank. This selection is based on factors such as the species of your original hybridoma and its known characteristics, aiming for optimal fusion efficiency and post-fusion stability.

02Expert Hybridoma Re-Fusion

Your existing hybridoma cells, which may be exhibiting instability or poor production, are expertly fused with the chosen myeloma partners. We employ advanced fusion protocols, including optimized polyethylene glycol (PEG)-mediated fusion techniques, to maximize the formation of viable hybrid-hybridoma cells.

03Rigorous Hybridoma-Myeloma Fusion Cell Screening

Following the fusion event, the cell mixture is cultivated in a highly selective medium (typically HAT medium). This critical step ensures that only successfully fused hybridoma-myeloma cells survive and proliferate, eliminating unfused parental myeloma cells and the original, unstable hybridoma cells.

04Identification of Positive Antibody-Producing Cell Lines

As new hybridoma colonies emerge and expand, their supernatants are meticulously screened to identify clones actively secreting your target antibody. This involves employing sensitive and specific assays, often tailored to your antibody, such as ELISA or Western Blotting, to confirm desired antibody production.

05Subcloning and Cryopreservation of Finalized Positive Cell Lines

Identified positive cell lines that demonstrate robust antibody production are then subjected to several rounds of limiting dilution subcloning. This essential step guarantees the monoclonality of the rescued line and allows for the selection of the most stable and high-yielding clones. Finalized top clones are expanded and cryopreserved to establish reliable cell banks for your future use.

Features of Our Service

Highly Customized Service Plans:

Recognizing that each hybridoma and research project is special, we provide individualized service options. These are meticulously designed based on the specific condition of your cell line and your intended experimental outcomes, ensuring a bespoke approach.

Meticulous Experimental Design and Execution:

Our scientific team employs rigorous experimental design principles. All procedures are conducted under strictly tested and optimized conditions within our state-of-the-art facilities, maximizing the potential for a successful rescue.

Exceptional Value for Your Investment:

We strive to deliver high-cost performance, providing a scientifically robust service that helps you save on your project budget by recovering valuable assets rather than re-developing them from scratch.

Efficient Project Turnaround:

Understanding the time-sensitive nature of research, we are dedicated to providing a short turnaround time for our re-fusion services, getting your rescued hybridomas back to you as efficiently as possible without compromising quality.

FAQ

Q: How do you select the myeloma fusion partners for my specific hybridoma?
A: We maintain a panel of well-characterized, high-performance myeloma cell lines. The selection process considers the species of origin of your hybridoma (e.g., mouse, rat), any available data on its previous fusion partner if known, and the growth characteristics desired. Using at least two different myeloma lines in parallel increases the chances of finding a highly compatible and efficient partner.
Q: My original hybridoma is in very poor condition (low viability, no detectable antibody). Is re-fusion still an option?
A: Even if your hybridoma is in poor condition, re-fusion may still be a viable option, provided some viable cells are remaining that harbor the genetic material for your antibody. The re-fusion process is designed to provide a new, healthier cellular environment. We recommend discussing the specifics of your cell line with our experts for a thorough evaluation.
Q: What if multiple stable, high-producing clones are rescued from the re-fusion? Do I get all of them?
A: Yes, if our re-fusion and subcloning efforts yield multiple distinct clones that meet the criteria for stability and high antibody production, we will discuss these findings with you. Typically, we will provide you with an agreed-upon number of the best-performing clones (e.g., top 2-3) as part of the standard service, with options to receive additional characterized clones if desired.
Q: How is the antibody production of the rescued clones confirmed and quantified?
A: Antibody production is initially confirmed through screening assays like ELISA or Western blot, using culture supernatants from emerging clones. For lead candidate clones, we can perform more quantitative assessments of antibody secretion rates (e.g., µg/ml/10^6 cells/day) to ensure they meet your productivity requirements. The specific assays can often be tailored using antigen or detection reagents you provide.
Q: What are the final deliverables I will receive after completing the service?
A: Upon completion, you will receive cryopreserved vials of the successfully rescued and subcloned hybridoma cell line(s) – typically 2-5 vials per selected clone. You will also receive a comprehensive final report detailing all methodologies, screening data, subcloning results, and characterization of the rescued clones, including viability and antibody productivity assessments.

Equipped with years of experience in hybridoma technology, along with novel tech platforms and an expert team, Creative Biolabs is capable of assisting you in your projects with reliable hybridoma rescue services including but not limited to hybridoma re-fusion. Please feel free to contact us for inquiries into your customized services.

Reference

Wu, Meng, et al. "Substantially improved electrofusion efficiency of hybridoma cells: Based on the combination of nanosecond and microsecond pulses." Bioengineering 9.9 (2022): 450.
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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