Antibodies, A Laboratory Manual, Second Edition, Edited by Edward A. Greenfield



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Preparing Antigens Using a Baculovirus Expression System

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Baculoviruses contain double-stranded, circular, supercoiled DNA in a rod-shaped capsid. The viral life cycle includes three major phases: (1) early phase or virus synthesis phase (0.56 h after infection), which includes viral attachment, uncoating early viral gene expression, and shutting off host gene expression; (2) late phase (612 h after infection), which includes viral replication and formation of budded virus containing host plasma membrane and glycoprotein (gp)64, which is required for virus entry; and (3) very late phase (2496 h after infection), which includes formation of polyhedral inclusion bodies and cell lysis. Major advantages of a baculovirus expression system are (1) safety, because baculoviruses are nonpathogenic to mammals and plants; (2) posttranslational modifications of the protein are similar to those of mammals; and (3) insect cells can be easily grown in suspension culture, which makes protein production easy to scale up.

Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition
Antibodies: A Laboratory Manual, Second edition

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