Recombinant Antibodies and In Vitro Selection Technologies

Geyer, C. Ronald; McCafferty, John; Dübel, Stefan; Bradbury, Andrew; Sidhu, Sachdev S.

doi:10.1007/978-1-61779-931-0_2

Cited by 65 publications

(42 citation statements)

References 178 publications

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“…Therefore, synthetic libraries of human 55 antibody fragments enable obtaining human antibodies that recognize human antigens, which is of 56 great importance for possible therapeutic applications. Following a selection of specific Fabs or 57 scFvs, further genetic modifications of their cDNAs which substitute for a natural process of 58 antibody maturation may result in the generation of high-affinity and high-specificity molecules [3]. 59…”

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confidence: 99%

“…Natural libraries of Fab or single chain variable fragments (scFv) 50 are increasingly being displaced by highly diverse semisynthetic or synthetic (artificial) ones. These 51 are constructed by introducing substantial randomizations of defined amino acids within CDRs of a 52 chosen, stable V H and/or V L framework using genetic engineering methods [2,3]. The major 53 advantage of such libraries is their independence from a natural repertoire of immunoglobulins, 54 which obviously lack self-antigen-specific antibodies [2].…”

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confidence: 99%

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A new expression vector facilitating production and functional analysis of scFv antibody fragments selected from Tomlinson I + J phagemid libraries

et al. 2015

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A new expression vector facilitating production and functional analysis of scFv antibody fragments selected from Tomlinson I + J phagemid libraries

et al. 2015

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“…Since the first description that antibody fragments can be expressed on the surface of filamentous bacteriophage and selected for specific binding to a target, phage display has become one of the most widely used in vitro approaches for the generation of antibodies for research and therapeutic applications [1,2]. More recently, the display of antibodies at the surface of yeast or mammalian cells, as well as cell free systems, such as ribosome display, have complemented the panel of technologies available for the isolation and engineering of antibodies and other protein scaffolds [3,4].…”

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confidence: 99%

Purpose-Oriented Antibody Libraries Incorporating Tailored CDR3 Sequences

et al. 2015

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Abstract:The development of in vitro antibody selection technologies has allowed overcoming some limitations inherent to the hybridoma technology. In most cases, large repertoires of antibody genes have been assembled to create highly diversified libraries allowing the isolation of antibodies recognizing virtually any antigen. However, these universal libraries might not allow the isolation of antibodies with specific structural properties or particular amino acid contents that are rarely found in natural repertoires. Purpose-oriented libraries specially designed to incorporate desired characteristics have been successfully used. However, the workload required for library construction has limited the attractiveness of this approach compared to the use of large universal libraries. We have developed an approach to capture synthetic or natural diversity into the complementarity determining regions 3 (CDR3) of human antibody repertoires using Type IIS restriction enzymes. In this way, we generated several libraries either biased in amino acid content or towards long CDRH3 loops. The latter were successfully used to identify antibodies inhibiting the enzymatic activity of horseradish peroxidase, whereas libraries enriched in histidines allowed for the isolation of antibodies binding to human Fc in a pH-dependent manner. These libraries indicate that tailored diversification of CDR3 is sufficient to generate purpose-oriented libraries and isolate antibodies with uncommon properties. OPEN ACCESSAntibodies 2015, 4 104

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“…Recombinant antibody technology enables the screening of large libraries to generate monoclonal antibodies in a shorter time than conventional methods [1]. Libraries with a vast repertoire and efficient screening are two prerequisites for obtaining monoclonal antibodies with high affinity to the antigen [2,3].…”

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Single-step Colony Assay with Autoinduction of scFv Expression for the Screening of Antibody Libraries

Kato¹,

Hanyu

2019

BioTechniques

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We present a simple colony assay method for screening antibody libraries based on autoinduction of antibody fragment expression. This protocol eliminates the need for colony size monitoring and a separate induction step for single-chain Fv (scFv) antibody fragment expression. Here, scFvs are expressed in an automatic and timely fashion during the assay, resulting in high yields of positive clones and substantial time savings. The method was used successfully to establish monoclonal scFvs with high affinity and specificity against human IgG.

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Recombinant Antibodies and In Vitro Selection Technologies

Cited by 65 publications

References 178 publications

A new expression vector facilitating production and functional analysis of scFv antibody fragments selected from Tomlinson I + J phagemid libraries

A new expression vector facilitating production and functional analysis of scFv antibody fragments selected from Tomlinson I + J phagemid libraries

Purpose-Oriented Antibody Libraries Incorporating Tailored CDR3 Sequences

Single-step Colony Assay with Autoinduction of scFv Expression for the Screening of Antibody Libraries

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