Isolation of two distinct populations of recombinant antibody molecules specific for rat malonyl-CoA decarboxylase from a semi-synthetic human scFv display library using Ex-phage system

Baek, Hyun Jung; Hur, Byung Woong; Cho, Jin Won; Lee, Hee Kyung; Kim, Namil; Oh, Moon Young; Hoon, Sang

doi:10.1016/j.imlet.2003.11.010

Cited by 3 publications

(2 citation statements)

References 20 publications

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“…In our approach, efficiency in the antigen-specific selection seemed to have increased at a large extent as in the case of Ex-phage in combination with the non-supE JS5 cells, which showed over a 100-fold increase in panning efficiency (Baek et al, 2002). However, a profound decrease in TU of the eluted phages after a successive round of panning was observed at the Ex-phage/TOP10F′ combination due to the absence of the wt pIII on affinity-selected phages (Baek et al, 2004), but this adverse phenomenon was not observed in the phage progenies from the Ex12/XL-1 Blue MRF′ combination. It is interesting to note that the phage progenies obtained from the Ex12/XL-1 Blue MRF′ combination showed a slightly higher display level of the L chain fragments and a 5-10 fold better antigen-binding functionality than those from the Ex12/TOP10F′ combination.…”

Section: Discussionmentioning

confidence: 97%

Enhancing phage display of antibody fragments using gIII-amber suppression

Joo

Hur

et al. 2007

Gene

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Section: Discussionmentioning

confidence: 97%

Enhancing phage display of antibody fragments using gIII-amber suppression

Joo

Hur

et al. 2007

Gene

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“…After four rounds of selection and amplification, the recoveries of phages through successive rounds of selection are shown in Table 1. The IL-21R-binding scFv-phages were enriched significantly after each round (Baek et al, 2004).…”

Section: Biopanning Of Il-21r-binding Scfvsmentioning

confidence: 99%

Expression and Characterization of Recombinant Interleukin-21 Receptor and Its Targeting Single-Chain Variable Fragment Antibodies Selected from a Human Phage Display Library

Zhang

Luo

et al. 2012

DNA and Cell Biology

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Interleukin-21 receptor (IL-21R) is widely expressed in lymphocytes, and plays an important role in immunological cell proliferation and cytokine production. The present study aims to express a recombinant extracellular domain of human IL-21R (rhIL-21R-ECD) with high yield, and to screen the anti-IL-21R single-chain variable fragments (scFvs) from a synthetic human phage display library. The rhIL-21R-ECD, being expressed mainly as insoluble inclusion bodies in Escherichia coli BL21 (DE3), was purified and refolded. ELISA analysis showed that the refolded rhIL-21R-ECD bound to its ligand IL-21 in a concentration-dependent manner. Using a phage display technique, anti-IL-21R scFvs were screened from a naïve human phage display library by biopanning. After four rounds of panning, positive clones were isolated, sequenced, and characterized. The clone with highest activity was designated as C2. Flow cytometry analysis showed that the scFv C2 could recognize IL-21R on Jurkat cells. Furthermore, proliferation assay revealed a concentration-dependent inhibitory effect of C2 on the Jurkat cell, with fifty percent inhibitory concentration (IC(50)) of 78 nM. A human scFv antibody C2 with a high binding specificity to IL-21R was isolated and characterized. The antibody showed a concentration-dependent inhibitory effect on Jurkat cell proliferation.

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Antibody‐based Proteomics: From bench to bedside

Dahan¹,

Chevet

Liu

et al. 2007

Proteomics Clinical Apps

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Over the past 75 years, antibodies have gone from being recognized as disease biomarkers to being used as very powerful therapeutic tools. This evolution has been accelerated by the identification of mAb and the extensive use of immunological tools both at fundamental and clinical levels. In this review, we evaluate how antibodies can be used to assess the proteome of cells or tissues and their relevance for clinical applications. These antibody-based proteomics approaches also require analytical and technological pipelines as well as specific enabling tools which are described. Our first objective was to establish how large-scale datasets (provided by high-throughput studies such as proteomics and transcriptomics) can be integrated with literature searches and clinical data to identify potentially relevant markers against which antibodies should be raised. Then based on an extensive literature review and our experience, we compare the methodologies developed to produce specific antibodies either in vivo or in vitro. This is followed by the description of the validation tools currently available and it also includes the use of antibody-based approaches in the establishment of molecular signatures utilized at the bench and soon available for bedside use.

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Isolation of two distinct populations of recombinant antibody molecules specific for rat malonyl-CoA decarboxylase from a semi-synthetic human scFv display library using Ex-phage system

Cited by 3 publications

References 20 publications

Enhancing phage display of antibody fragments using gIII-amber suppression

Enhancing phage display of antibody fragments using gIII-amber suppression

Expression and Characterization of Recombinant Interleukin-21 Receptor and Its Targeting Single-Chain Variable Fragment Antibodies Selected from a Human Phage Display Library

Antibody‐based Proteomics: From bench to bedside

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