Hybrido Hybridoma Cloning Factor

R13-HB01-1L

Hybrido Hybridoma cloning factor is the first completely defined growth promoting supplement especially for hybridoma cells. It may substantially increase the cloning efficiency of the newly fused hybridoma cells.

Introduction
  Hybridoma cells, which are used for production of monoclonal antibodies, result from fusion of an antibody producing B-cell with a tumor cell, typically myeloma cells. Growth factors and serum are generally used to support the hybridoma development and achieve an optimal cell density and cloning efficiency.
  Hybrido Hybridoma cloning factor is the first completely defined growth promoting supplement especially for hybridoma cells. It may substantially increase the cloning efficiency of the newly fused hybridoma cells. It also significantly increases the successful rate of monoclonization.
  In comparison with the method of using feeder layer cells for supporting the growth of hybridoma cells, Hybrido Hybridoma cloning factor is not only more effective but provides numerous advantages.

Advantages
*Increase the cloning efficiency of the newly fused hybridoma cells
*Increase the successful rate of monoclonization


The recommend Timing of adding Hybrido Cloning Factor
1. Cell culture media for the monoclonization of hybridoma cells.
2. HAT-media during the selection.


Experiment Data

A significant higher cloning efficiency was observed for the usage of Hybrido

Figure 1. A significant higher cloning efficiency was observed for the usage of Hybrido.
Comparison of the cloning efficiency of the newly fused hybridoma cells. The newly PEG
fused hybridoma cells were plated into a 96-well plate containing FCS media with
Hybrido (green bars), FCS media with feeder layer (blue bars), or regular FCS media
(white bars). Hybridoma cells were subject to HAT selection 14 days after the cell fusion. The
numbers of viable hybridoma colonies were visually counted under a microscope. Two mouse
myeloma fusion partners, NS-1 and SP2/0, were evaluated by four independent fusion
experiments with freshly prepared mouse spleens.




Figure 2: A significant higher successful rate of monoclonization was observed for the
usage of Hybrido.
Comparison of successful rate of monoclonizing hybridoma cells. Eight
clones of hybridoma cells from NS-1 or SP2/0 fusion partners were monoclonized in the
media containing FCS media with Hybrido (green bars), FCS media with feeder layer (blue
bars), or regular FCS media (white bars). The numbers of viable hybridoma colonies in a well
were visually counted under a microscope.



Figure 3. The usage of Hybrido will not affect the yield of secreting Ab in a hybridoma cell.
The data shows the result of comparison of the titers of secreting Ab.A clone of hybridoma cells
(anti human transferrin, L3B5) was cultured in the media containing FCS media with Hybrido (green
bars), FCS media with feeder layer (blue bars), or regular FCS media (white bars) for seven days.
The supernatants were harvest and examined by the titer of secreting Ab by ELSIA assay.


Q & A

1. Can I add Hybrido Cloning Factor for sustaining the growth of hybridoma cells?
Yes, we also suggest routinely supplying Hybrido Cloning Factor to culture the established
hybridoma cells.  Hybrido Cloning Factor may promote the growth of hybridoma cells.


2. Once using Hybrido Cloning Factor to sustain hybridoma cells, does hybridoma cells
comprise a growth dependent phenomenon or would cells die if stop using Hybrido
Cloning Factor?

No, Hybrido Cloning Factor is a chemical defined growth promoting supplement. It will not
cause a growth dependent phenomena in hybridoma cells. Furthermore, Hybrido Cloning
Factor does not affect the secretion of antibody in hybridoma cells, either.


Technical Datasheet