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From: "Max Warncke" <max.warncke@uniklinik-freiburg.de>
To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
Subject: Summary to Factors enhancing hybridoma secretion
Date: Fri, 28 Jan 2005 16:31:45 +0100
Thank you very much for all the intersting anwser to my question:
Dear Flowers (and antibody producers),
I want to isolate the idiotype of the BALB/c lymphoma A20 from
cell culture supernatant. As A20 is not secreting much I was
wondering, if i could add factors which enhance secretion in
Hybridoma. Do you know any cytokines or reagents, which could
stimulate the B cell to secrete more of its idiotype.
Summary:
“Your best bet is to clone the cells, 1 cell per well (round bottom plate) by FACS (duh) and screen for the best producer. Also you can grow the cells in 20% FBS.
I used to use a growth factor as well.”
“You can make hybridomas of A20 with a fusing myeloma which has some other selectable gene such as Neo”
“We use Hybridoma Cloning Factor (Bioveris) but it mainly helps to allow hybridomas to grow out from single clones, not really know whether it will boost production. Scaleup is the other option”
“Try IL-6. That seems to be working for us. Another option is to use culture supes from PHA-stimulated thymocytes.”
“Gibco/Invitrogen sells a product called Optimab, supposedly increasing secretion by twofold or so.”
From BD: “These guys could try the BD Cell MAb medium with serum supplementation. In general you see anywhere from 3-10 fold yield increase in batch production when using aerated systems such as spinner, rollers, shakers, etc. Catalogue number is 220511.”
Max Warncke, Dipl. Biol.
University Freiburg Medical Center
Medicin I
AG Veelken
Lab. Nothnagel / 5.OG
Breisacherstr. 117
79106 Freiburg
fon: +49.761.270- 7196
fax: +49.761.270- 7177
