Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2005-05-23
2009-12-01
Liu, Sue (Department: 1639)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S005000, C435S069700, C435S320100, C536S023100, C536S023500
Reexamination Certificate
active
07625700
ABSTRACT:
The present invention describes a rapid and efficient in vivo library-versus-library screening strategy for identifying optimally interacting pairs of heterodimerizing polypeptides. It allows for the screening of a protein library against a second protein library and therefore finds numerous applications in the study of protein-protein interactions. Two leucine zipper libraries, semi-randomized at the positions adjacent to the hydrophobic core, were genetically fused to either one of two designed fragments of the enzyme murine dihydrofolate reductase (mDHFR), and cotransformed intoE. coli. Interaction between the library polypeptides was required for reconstitution of the enzymatic activity of mDHFR, allowing bacterial growth. Using more weakly associating mDHFR fragments, we increased the stringency of selection. We applied these selection processes to a library-versus-library sample of 2.0×106combinations, and selected a novel leucine zipper pair which may be appropriate for use in further in vivo heterodimerization strategies.
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Arndt Katja M.
Michnick Stephen William Watson
Pelletier Joelle N
Pluckthun Andreas
Angres Issac A.
Liu Sue
Odyssey Thera Inc.
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