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Antibody Production & Purification

Reduce Hybridoma Screening & Cloning Time by 75%




Rapid Generation of Monoclonal Antibodies

Monoclonal antibodies (mAb) are powerful tools which are widely used in every phase of the biomedical field from discovery research and diagnosis to therapeutics. Yet, existing methods for isolating mAb-producing hybridomas are tedious and time consuming and often not amenable to high-throughput screening. In particular, the most challenging, time-consuming, and labor-intensive phase in generating mAbs is the Limiting Dilution Subcloning (LDS) step. During this step, fused hybridoma cells are added to microtiter plate wells containing media supplemented with HAT (hypoxanthine, aminopterin, thymidine). Following selection, hyridbomas are screened for reactivity (using methods such as ELISA) and processed through cycles of LDS until a positive cell is found to be clonal. Prior attempts to improve the process have lacked efficiency and are often cost prohibitive.

 

A Novel Technology for mAb Detection and Isolation

The Direct Selection of Hybridomas (DiSH) kit is a novel system in which mAb-secreting hybridoma cells co-express significant amounts of the membrane form of the secreted immunoglobulin (Ig) on their surfaces. Subsequently, hybridomas with high-affinity for the target antigen are labeled. And once labeled, these hybridomas can be isolated from among a pool of hundreds of thousands of cells in a matter of hours using Fluorescence activated cell sorting (FACS) or magnetic separation. The total elapsed time from harvest to the first screening is typically 16 days, resulting in improved efficiency for detection of highly specific mAb.

Key Features of the Technology:

  • Decreased time and labor costs
  • Improved selection of specific, stable hybridomas
  • Minimal manipulation of cloned cell lines relative to approaches requiring Limiting Dilution Subcloning

Application Note

Download the Application Note: Anti-hESC mAb Hesca-2 Binds to a Glycan Epitope Commonly Found on Carcinomas.

 

Create Mouse Hybridoma

Introducing two new cloning media, ABEOCLONE™ Base Semi-solid and CHO TCSC Semi-solid media, for creating stable mouse hybridomas or CHO cell lines.

  • Compared with limited dilution cloning, semi-solid cloning is faster and requires fewer resources to isolate monoclonal cell lines
  • Enables isolation of diverse clones with a variety of growth rates and productivities compared to selection in bulk liquid medium
  • Rare, high-producing clones are easily isolated compared with selection in liquid medium


Sp2ab myeloma fusion partner 2nd-ary Abs DiSH%trade; Kit

Create Stable CHO Cell Line


AbeoClone™ (CHO TCSC Semi-solid Medium) Nuclear-ID® Kits & Dyes Cell Viability Kits & Dyes

How Does the DiSH™ Technology Work?

The isolation of antigen-specific hybridomas is performed through a four-step procedure.

  1. IMMUNIZE: Immunization of the mouse with target antigen
  2. FUSE: B cells are fused with Abeome’s Igα and Igβ-expressing myeloma, Sp2ab, resulting in hybridomas that secrete and surface express their specific immunoglobulin
  3. LABEL/ENRICH: Antigens are conjugated to biotin to enable capture of antigen-specific hybridomas. Hybridomas can be fluorescently labeled for use with FACs or separated by magnetic beads.
  4. CLONE: Antigen-specific hybridomas are subsequently isolated and cloned by:
    1. FACS: single cell suspensions are incubated and resulting supernatant analyzed for secreted antibody, or
    2. Magnetic bead separation: cells plated into AbeoClone™ Medium (semi-solid agar) are examined for the presence of visible colonies.
 

DiSH™ Kit 

Reduce labor time and costs using a novel technology for rapid hybridoma production
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ENZ-71001-0001 1 Kit 1,050.00 USD
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Sp2ab myeloma fusion partner 

Myeloma fusion partner for rapid hybridoma production
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ENZ-70008-0001 1 Vial 945.00 USD
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AbeoClone™ (HAT Semi-solid Medium) 

Semi-solid HAT media optimized for selection of hybridoma colonies
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ENZ-70004-0090 90 ml 314.00 USD
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AbeoClone™ (Base Semi-solid Medium) (2X) 

Semi-solid Base media optimized for selection of hybridoma colonies and monoclonal cell line isolation.
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ENZ-70009-0045 45 ml 152.00 USD
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AbeoClone™ (CHO TCSC Semi-solid Medium) 

Semi-solid CHO TCSC media optimized for monoclonal cell line isolation and development.
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ENZ-70010-0090 90 ml 305.00 USD
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PCM (Prefusion Medium) 

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ENZ-70001-0200 200 ml 62.00 USD
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WCM (Wash Medium) 

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ENZ-70002-0200 200 ml 53.00 USD
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FCM (Fusion Medium) 

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ENZ-70003-0200 200 ml 62.00 USD
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RCM (Recovery Medium) 

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ENZ-70005-0200 200 ml 314.00 USD
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ECM (Expansion Medium) 

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ENZ-70006-0200 200 ml 104.00 USD
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PEG (polyethylene glycol) 

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ENZ-70007-0001 1.5 ml 53.00 USD
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