Engineering resistance to trypsin inactivation into L-asparaginase through the production of a chimeric protein between the enzyme and a protective single-chain antibody.
We have demonstrated that a trypsin sensitive enzyme such as L-asparaginase can be rendered trypsin resistant by genetically fusing its gene with that of a single-chain antibody derived from a preselected monoclonal antibody capable of providing protection against trypsin. The chimeric L-asparaginase retained 75% of its original activity upon exposure to trypsin, whereas the native unprotected L-asparaginase control was totally inactivated.
Newsted
W J
WJ
Department of Molecular Biology and Genetics, University of Guelph, Ontario, Canada.
Ramjeesingh
M
M
Zywulko
M
M
Rothstein
S J
SJ
Shami
E Y
EY
D000911:Antibodies, Monoclonal / Q000235:genetics; D001215:Asparaginase / Q000187:drug effects / Q000235:genetics*; D001483:Base Sequence; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000187:drug effects; D014357:Trypsin / Q000276:immunology / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D014357:Trypsin; D001215:Asparaginase