Hybridomas generated from secretory plasmocytes do not have surface Igs. It is therefore impossible to carry out a selective sorting by FACS because the secreted Igs will only have a furtive attachment to the plasma membrane.
There are, however, methods for trapping the antibodies on the surface of the secretory cell, using a polymer which coats the cell and entraps the antibodies on the surface of the hybridoma.
Another method uses bispecific antibodies capable of binding to the surface of the hybridoma and capturing the antibody at the outlet of the cell.
But this remains expensive and delicate methods, which can not replace the usual techniques, except in extreme cases where the hybridoma is unstable and under represented.
Thank you Dr. Pin, for your very informative reply.
Indeed, the task of flow-sorting hybridoma based on surface IgG expression has been met with several challenges. However, my experience was limited to the anti-AChR (mAb35) cell-line TIB-175.
The most critical limitation was that cells stained with anti-IgG (minimal x-reactivity) experienced very low viability (~5%) post-sort even when the conditions of the sorting were 4 °C and suspended in PBS/0.5% BSA (absence of heat and complement). What made me conclude that this is a result of antibody engagement is that the negatively-sorted population did not experience a decrease in viability. The possibility that this result was due to the antibody type or clone I am using was also eliminated by having another group report similar results with a different antibody.
Your point about hybridoma generated from plasmocytes is new to me and I still need to research it further. Could there by hybridomas generated from cells other than plasma cells? TIB-175 was generated by hybridizing a rat B cell and a mouse myeloma.
Thank you very much for your advice.
Perhaps this publication might be interesting for you:
https://www.ncbi.nlm.nih.gov/pubmed/25058593
Hi Dr. Weller,
Thank you very much for attaching this useful publication.
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