2021
DOI: 10.1002/cpz1.182
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Generation of Single‐Chain Variable Fragment (scFv) Libraries for Use in Phage Display

Abstract: Phage display is a powerful platform for the discovery of novel biologics with high binding affinities to a specific target protein. Here, we describe methods to construct a phage display library containing diverse single-chain variable antibody fragments (scFvs). Specifically, updated methods for polymerase chain reaction (PCR) amplification and fusion of human antibody genes, their ligation into the pComb3X vector for transformation into 5αF I q competent bacterial cells, and their expression in M13KO7 helpe… Show more

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Cited by 4 publications
(3 citation statements)
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“…Modern recombinant molecular biology techniques have simplified the creation of highly diverse libraries that reproduce naturally occurring binder repertoires (see Current Protocols article: Schladetsch & Wiemer, 2021) that expand on protein folds used for cellular protein-protein interactions. An example of the latter is ankyrin repeat proteins, which are naturally occurring proteins composed of repeating 33 amino acid modules, each consisting of a β-turn followed by two anti-parallel α-helices (Fig.…”
Section: Introductionmentioning
confidence: 99%
“…Modern recombinant molecular biology techniques have simplified the creation of highly diverse libraries that reproduce naturally occurring binder repertoires (see Current Protocols article: Schladetsch & Wiemer, 2021) that expand on protein folds used for cellular protein-protein interactions. An example of the latter is ankyrin repeat proteins, which are naturally occurring proteins composed of repeating 33 amino acid modules, each consisting of a β-turn followed by two anti-parallel α-helices (Fig.…”
Section: Introductionmentioning
confidence: 99%
“…Nevertheless, this technique has limitations. For instance, most mAbs produced via hybridoma technology are of murine origin and may induce human anti-mouse antibody responses in clinical settings [14]. To mitigate immunogenicity, antibodies from non-human species undergo a humanization process, either through chimerization or by grafting complementarity-determining regions (CDRs) on scaffolds from human Ig.…”
Section: Introductionmentioning
confidence: 99%
“…This study would provide valuable guidance for selecting alternative antibiotics by conducting diagnostic tests with alternative β-lactams. It is essential to consider that the antigenic determinant represents a critical component in drug allergy testing, and it accounts for the discrepancies observed among these tests. , We propose the production of a synthetic IgE molecule as a viable alternative. By constructing an immune combinatorial human scFv library and using phage display technology, we selected scFv variants with high affinity and selectivity for amoxicillin.…”
Section: Introductionmentioning
confidence: 99%