An efficient method for isolating antibody fragments against small peptides by antibody phage display

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An efficient method for isolating antibody fragments against small peptides by antibody phage display. / Duan, Zhi; Siegumfeldt, Henrik.

In: Combinatorial Chemistry & High Throughput Screening, Vol. 13, 2010, p. 818-828.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Duan, Z & Siegumfeldt, H 2010, 'An efficient method for isolating antibody fragments against small peptides by antibody phage display', Combinatorial Chemistry & High Throughput Screening, vol. 13, pp. 818-828.

APA

Duan, Z., & Siegumfeldt, H. (2010). An efficient method for isolating antibody fragments against small peptides by antibody phage display. Combinatorial Chemistry & High Throughput Screening, 13, 818-828.

Vancouver

Duan Z, Siegumfeldt H. An efficient method for isolating antibody fragments against small peptides by antibody phage display. Combinatorial Chemistry & High Throughput Screening. 2010;13:818-828.

Author

Duan, Zhi ; Siegumfeldt, Henrik. / An efficient method for isolating antibody fragments against small peptides by antibody phage display. In: Combinatorial Chemistry & High Throughput Screening. 2010 ; Vol. 13. pp. 818-828.

Bibtex

@article{886370cc60734be293ddddd4007d30e1,
title = "An efficient method for isolating antibody fragments against small peptides by antibody phage display",
abstract = "We generated monoclonal scFv (single chain variable fragment) antibodies from an antibody phage display library towards three small synthetic peptides derived from the sequence of s1-casein. Key difficulties for selection of scFv-phages against small peptides were addressed. Small peptides do not always bind efficiently to passive adsorption surfaces, and we developed a simple method to quantify the binding capacity of surfaces with the peptides. Background binding (the binding of scFvs to the background matrix) is an obstacle for successful selection, and we evaluated two methods that drastically reduced the background binding. An optimized method therefore enabled a panning procedure where the specific (peptide binding) scFv-phages were always dominant. Using 15-mer peptides immobilized on Nunc Immobilizer Streptavidin plates, we successfully generated scFvs specifically against them. The scFvs were sequenced and characterized, and specificity was characterized by ELISA. The methods developed in this study are universally applicable for antibody phage display to efficiently produce antibody fragments against small peptides.",
keywords = "LIFE, Phage display, scFv, Tomlinson I + J libraries, casein, peptide",
author = "Zhi Duan and Henrik Siegumfeldt",
year = "2010",
language = "English",
volume = "13",
pages = "818--828",
journal = "Combinatorial Chemistry & High Throughput Screening",
issn = "1386-2073",
publisher = "Bentham Science Publishers",

}

RIS

TY - JOUR

T1 - An efficient method for isolating antibody fragments against small peptides by antibody phage display

AU - Duan, Zhi

AU - Siegumfeldt, Henrik

PY - 2010

Y1 - 2010

N2 - We generated monoclonal scFv (single chain variable fragment) antibodies from an antibody phage display library towards three small synthetic peptides derived from the sequence of s1-casein. Key difficulties for selection of scFv-phages against small peptides were addressed. Small peptides do not always bind efficiently to passive adsorption surfaces, and we developed a simple method to quantify the binding capacity of surfaces with the peptides. Background binding (the binding of scFvs to the background matrix) is an obstacle for successful selection, and we evaluated two methods that drastically reduced the background binding. An optimized method therefore enabled a panning procedure where the specific (peptide binding) scFv-phages were always dominant. Using 15-mer peptides immobilized on Nunc Immobilizer Streptavidin plates, we successfully generated scFvs specifically against them. The scFvs were sequenced and characterized, and specificity was characterized by ELISA. The methods developed in this study are universally applicable for antibody phage display to efficiently produce antibody fragments against small peptides.

AB - We generated monoclonal scFv (single chain variable fragment) antibodies from an antibody phage display library towards three small synthetic peptides derived from the sequence of s1-casein. Key difficulties for selection of scFv-phages against small peptides were addressed. Small peptides do not always bind efficiently to passive adsorption surfaces, and we developed a simple method to quantify the binding capacity of surfaces with the peptides. Background binding (the binding of scFvs to the background matrix) is an obstacle for successful selection, and we evaluated two methods that drastically reduced the background binding. An optimized method therefore enabled a panning procedure where the specific (peptide binding) scFv-phages were always dominant. Using 15-mer peptides immobilized on Nunc Immobilizer Streptavidin plates, we successfully generated scFvs specifically against them. The scFvs were sequenced and characterized, and specificity was characterized by ELISA. The methods developed in this study are universally applicable for antibody phage display to efficiently produce antibody fragments against small peptides.

KW - LIFE

KW - Phage display

KW - scFv

KW - Tomlinson I + J libraries

KW - casein

KW - peptide

M3 - Journal article

VL - 13

SP - 818

EP - 828

JO - Combinatorial Chemistry & High Throughput Screening

JF - Combinatorial Chemistry & High Throughput Screening

SN - 1386-2073

ER -

ID: 32447964