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Combination of phag...
Combination of phage and Gram-positive bacterial display of human antibody repertoires enables isolation of functional high affinity binders
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- Hu, Francis Jingxin, 1986- (author)
- KTH Royal Institute of Technology,KTH,Proteomik och nanobioteknologi
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- Volk, Anna-Luisa (author)
- KTH Royal Institute of Technology,KTH,Proteomik och nanobioteknologi
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- Persson, Helena (author)
- KTH Royal Institute of Technology,Lunds universitet,KTH,Proteinteknologi,Science for Life Laboratory, SciLifeLab,Lund University, Sweden,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH
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- SÄLL, ANNA (author)
- Lund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH
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- Borrebaeck, Carl (author)
- Lund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH
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- Uhlén, Mathias (author)
- KTH Royal Institute of Technology,KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab,Technical University of Denmark
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- Rockberg, Johan (author)
- KTH Royal Institute of Technology,KTH,Proteomik och nanobioteknologi
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(creator_code:org_t)
- Elsevier, 2018
- 2018
- English.
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In: New Biotechnology. - : Elsevier. - 1871-6784 .- 1876-4347. ; 45, s. 80-88
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Abstract
Subject headings
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- Surface display couples genotype with a surface exposed phenotype and thereby allows screening of gene-encoded protein libraries for desired characteristics. Of the various display systems available, phage display is by far the most popular, mainly thanks to its ability to harbour large size libraries. Here, we describe the first use of a Gram-positive bacterial host for display of a library of human antibody genes which, when combined with phage display, provides ease of use for screening, sorting and ranking by flow cytometry. We demonstrate the utility of this method by identifying low nanomolar affinity scFv fragments towards human epidermal growth factor receptor 2 (HER2). The ranking and performance of the scFv isolated by flow sorting in surface-immobilised form was retained when expressed as soluble scFv and analysed by biolayer interferometry, as well as after expression as full-length antibodies in mammalian cells. We also demonstrate the possibility of using Gram-positive bacterial display to directly improve the affinity of the identified binders via an affinity maturation step using random mutagenesis and flow sorting. This combined approach has the potential for a more complete scan of the antibody repertoire and for affinity maturation of human antibody formats.
Subject headings
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Immunologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Immunology in the medical area (hsv//eng)
Keyword
- Affinity maturation
- Antibody
- Cell-surface display
- Flow cytometry
- HER2
- Phage display
- S. carnosus
- Binders
- Bins
- Cell membranes
- Display devices
- Genes
- Libraries
- Mammals
- Cell surface displays
- Antibodies
- Affinity maturation
- Antibody
- Cell-surface display
- Flow cytometry
- HER2
- Phage display
- S. carnosus
Publication and Content Type
- ref (subject category)
- art (subject category)
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