Post in B cell sequencing and antibody expression

Sabin,

Unfortunately we do not do single B cell cloning.  Hopefully someone on this
discussion forum does ... but in the mean time let's think this through
together.

So when you say you are having problems expressing functional antibodies from
the sorted B cells, what exactly do you mean?  Are you able to clone out and
express antibody but none are specific for the target?  If so, have you done a
control where you take half the spleen, fuse it and find the antigen specific
MAb in the traditionally way and in parallel do you single B cell cloning
approach.  If you do not get them with either approach, you know it was the
source of B cells.  If you get them with the traditional hybridomas, it is the
single B cell cloning protocol.

Frances

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On 11/1/21, 1:39 PM, "Antibody Discussion Forum on behalf of Sabin.Antony"
<email obscured> on behalf of <email obscured>> wrote:

    Hi I had some issues expressing  functional  antibodies from  target sorted
B cells. We expressed antibody sequences obtained by both single cell
sequencing and bulk B cell sequencing. I was expecting ~80% successful rate but
not much luck so far.  Any suggestion you can offer will be really helpful.
Thanks
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