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Abstract
Background In the therapeutic antibody development process, the yeast display technology which expresses a large library of antibodies is very useful for increasing the affinity of a lead antibody. Ideally, a yeast library should exceed the size of 10E10 to 10E11 to get close to the real affinity maturation process. However, due to low transformation efficiency with yeast, it requires trememdous scaling-up efforts to simply reach the 10E9 library size.
Methods To address the transformation problem, we developed a new electroporation device that applies a high voltage on a sealed electroporation tube containing the yeast and plasmids in a low conductance buffer.
Results The new device is arcing free due to the sealed design and each single reaction could generate 10E8 library size, far exceeding the 10E6 size that was previously reported in a single reaction.
Conclusions With the improved transformation efficiency, it becomes very straightforward to reach the currently difficult size of 10E9. Further more, it is possible to reach the 10E10 to 10E11 library size with reaction scaling-up. Our new method could be very useful for the field of antibody development.
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