title: Expression of single-domain antibodies in bacterial systems.
abstract: In this chapter we describe in detail the current protocols that are used to express single-domain antibodies in bacteria. Bacteria are among the most common expression systems for expressing recombinant proteins. We present different approaches for carrying out periplasmic and cytoplasmic expression, as well as small-scale and large-scale expression. In addition, we discuss the advantages and possible drawbacks of each protocol. We present data related to expression vectors, expression conditions, methods of protein extraction and purification, and yield and purity analysis of sdAbs. We also highlight important points that need to be considered before sdAbs that have been expressed in bacteria are used either in vitro or in vivo.
authors.affiliation: Institute for Biological Sciences, National Research Council of Canada, Ottawa, ON, Canada.
authors.forename: Toya Nath
authors.initials: TN
authors.lastname: Baral
authors.forename: Mehdi
authors.initials: M
authors.lastname: Arbabi-Ghahroudi
mesh_terms: D000915:Antibody Affinity / Q000276:immunology; D001419:Bacteria / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D061905:Single-Domain Antibodies / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies