hybridoma surface Ig expression

Joel Schildbach joel at ROSA.MIT.EDU
Thu Mar 25 15:28:51 EST 1993


F Garbrecht wrote:

>Do murine antibody producing hybridomas express any membrane bound
>antibody, or is all of it secreted?

Murine hybridomas can express membrane-bound antibody.  David
Parks and Leonard Herzenberg have taken advantage of this Ig
surface expression to select hybridoma variants with altered
hapten recognition.  In two-color FACS experiments, they determined
the ratio of labeled anti-mouse-Ig bound by a hybridoma (a measure
of surface Ig expression) to the amount of labeled hapten-
protein conjugate bound by a hybridoma.  By then selecting for
hybridomas with ratios of surface Ig to bound hapten that differ
significantly from a population of hybridoma cells, they could
isolate variants with altered hapten recognition.  For a more
coherent description, see Panka et al., (1988) PNAS, 85:3080-3084,

or Schildbach et al., (1991) J. Biol. Chem., 266:4640-4647.


Joel Schildbach
Dept. of Biology
Massachusetts Institute of Technology

joel at rosa.mit.edu





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