Expression of a heterodimeric Fab antibody protein in one cloning step

A new method is presented for creating antibody expression libraries in Escherichia coli. Rather than perform two cloning steps to express the heavy and light chains of the antigen binding domain, we have used a fusion-PCR method to link the coding regions for heavy and light chain sequences in a si...

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Bibliographic Details
Published in:	BioTechniques Vol. 12; no. 6; p. 864
Main Authors:	Mullinax, R L, Gross, E A, Hay, B N, Amberg, J R, Kubitz, M M, Sorge, J A
Format:	Journal Article
Language:	English
Published:	England 01-06-1992
Subjects:	Amino Acid Sequence Base Sequence Child, Preschool Cloning, Molecular - methods DNA Escherichia coli Genomic Library Humans Immunoglobulin Fab Fragments - genetics Molecular Sequence Data Polymerase Chain Reaction - methods
Online Access:	Get more information
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Summary:	A new method is presented for creating antibody expression libraries in Escherichia coli. Rather than perform two cloning steps to express the heavy and light chains of the antigen binding domain, we have used a fusion-PCR method to link the coding regions for heavy and light chain sequences in a single DNA molecule prior to vector ligation. This greatly simplifies the construction of antibody expression clones.
ISSN:	0736-6205