Construction of a large phage display antibody library by in vitro package and in vivo recombination

Cen, Xiaodong; Bi, Qun; Zhu, Shenggeng

doi:10.1007/s00253-006-0334-5

Cited by 12 publications

(4 citation statements)

References 18 publications

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“…99 This strategy increased the transformation efficiency and reduces the influences on diversity of library, which was caused by low conversion efficiency. 99 …”

Section: Future Perspectivesmentioning

confidence: 99%

Phagemid Vectors for Phage Display: Properties, Characteristics and Construction

Qiu

et al. 2012

Journal of Molecular Biology

132

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“…99 This strategy increased the transformation efficiency and reduces the influences on diversity of library, which was caused by low conversion efficiency. 99 …”

Section: Future Perspectivesmentioning

confidence: 99%

Phagemid Vectors for Phage Display: Properties, Characteristics and Construction

Qiu

et al. 2012

Journal of Molecular Biology

132

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“…Libraries generated using the traditional method generally contain 10 7 members, a size which is not sufficient enough for screening. Optimized and novel strategies to generate large antibody libraries have been carried out, increasing the diversity to 10 12 (23,39,121,126). However, both monovalently and polyvalently displayed scFvs and Fabs showed limitations that make them difficult for the selections based on intrinsic affinity (103).…”

Section: Phage and Phage Displaymentioning

confidence: 99%

Development of Anti-Infectives Using Phage Display: Biological Agents against Bacteria, Viruses, and Parasites

Huang

Bishop-Hurley

Cooper

2012

Antimicrob Agents Chemother

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The vast majority of anti-infective therapeutics on the market or in development are small molecules; however, there is now a nascent pipeline of biological agents in development. Until recently, phage display technologies were used mainly to produce monoclonal antibodies (MAbs) targeted against cancer or inflammatory disease targets. Patent disputes impeded broad use of these methods and contributed to the dearth of candidates in the clinic during the 1990s. Today, however, phage display is recognized as a powerful tool for selecting novel peptides and antibodies that can bind to a wide range of antigens, ranging from whole cells to proteins and lipid targets. In this review, we highlight research that exploits phage display technology as a means of discovering novel therapeutics against infectious diseases, with a focus on antimicrobial peptides and antibodies in clinical or preclinical development. We discuss the different strategies and methods used to derive, select, and develop anti-infectives from phage display libraries and then highlight case studies of drug candidates in the process of development and commercialization. Advances in screening, manufacturing, and humanization technologies now mean that phage display can make a significant contribution in the fight against clinically important pathogens.

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“…These studies showed combinatorial L chain variability alone is capable of generating a complex antibody repertoire. In another study, induction of further V H and V L re-assortment with Cre recombinase in a naive scFv library increased its calculated theoretical complexity from 10 7 to 10 11 (Sblattero and Bradbury, 2000; Cen et al, 2006). When this library was used in selections for 18 recombinant targets, an average of 6 specific antibodies were obtained per target.…”

Section: Discussionmentioning

confidence: 99%

Identification of recombinant antibodies against multiple distinct toll-like receptors by homolog mining a single immune scFv phage library

Chen¹,

Lipes²,

Kenan³

et al. 2009

Journal of Immunological Methods

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The generation of recombinant single-chain antibodies from either non-immune or immune phage display antibody libraries is an effective means to obtain high affinity antibodies against a specific target. Non-immune libraries contain a wide variety of antibodies but these are often low affinity. Immune libraries contain a high frequency of high affinity antibodies, but are typically limited to a single antigen. Due to the VH and VL recombination that occurs during antibody library construction, we hypothesized that an immune antibody library produced against one member of a protein family would contain antibodies specific for other members of the same protein family. Here, we tested this hypothesis by mining an existing anti-human Toll-like receptor-2 (hTLR2) antibody library for antibodies specific for other members of the TLR family. This procedure, referred to as homolog mining, proved to be effective. Using a cell-based system to pan and screen the anti-TLR2 library, we identified single chain antibodies specific for three of the four hTLR2 homologs we targeted. The antibodies identified, anti-murine TLR2, anti-hTLR5, and anti-hTLR6, bind specifically to their target, with no cross-reactivity to hTLR2 or other TLRs tested. These results demonstrate that combinatorial re-assortment of VH and VL fragments from multiple sources during Ab library construction increases Ab repertoire complexity, allowing antibody libraries produced by immunization with one antigen to be used to obtain antibodies specific to related antigens. The principle of homolog mining may be extended to other protein families and will facilitate and accelerate antibody production processes.

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Construction of a large phage display antibody library by in vitro package and in vivo recombination

Cited by 12 publications

References 18 publications

Phagemid Vectors for Phage Display: Properties, Characteristics and Construction

Phagemid Vectors for Phage Display: Properties, Characteristics and Construction

Development of Anti-Infectives Using Phage Display: Biological Agents against Bacteria, Viruses, and Parasites

Identification of recombinant antibodies against multiple distinct toll-like receptors by homolog mining a single immune scFv phage library

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