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Engineering the assembly pathway of the baculovirus-insect cell expression system
T A Hsu1, J J Eiden, M J Betenbaugh
1Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218.
Annals of the New York Academy of Sciences
|May 2, 1994
Summary
Researchers engineered insect cells to improve the production of functional antibodies. By introducing genes for immunoglobulin binding protein (BiP) and protein disulfide isomerase (PDI), they reduced abnormal protein aggregates.
Area of Science:
- Biotechnology
- Molecular Biology
- Protein Engineering
Background:
- Recombinant DNA technology enables antibody synthesis, but expression in baculovirus-insect cell systems yields functional antibodies alongside abnormal aggregates.
- Abnormal protein complex formation may stem from low intracellular levels of immunoglobulin binding protein (BiP) and protein disulfide isomerase (PDI).
Purpose of the Study:
- To engineer the baculovirus-insect cell expression system to enhance the production of functional antibodies.
- To investigate the role of BiP and PDI in antibody assembly and aggregation within insect cells.
Main Methods:
- Genes for murine BiP and PDI were cloned into the baculovirus vector (Autographa californica nuclear polyhedrosis virus) to create AcBB-BiP and AcBB-PDI.
- Spodoptera frugiperda (Sf-9) insect cells were infected with these recombinant baculoviruses.
Main Results:
- Infected Sf-9 cells produced recombinant BiP and PDI proteins of the correct molecular weight.
- These recombinant proteins were recognized by specific antibodies, confirming their successful expression.
Conclusions:
- Cloning BiP and PDI genes into the baculovirus vector is a viable strategy for engineering protein assembly pathways.
- This approach has the potential to reduce protein aggregation and increase the yield of active antibody oligomers.