TY - JOUR

T1 - Isolation of recombinant phage-displayed antibodies recognizing skin keratinocytes

AU - Jensen, Kim Bak

AU - Kristensen, Peter

PY - 2005

Y1 - 2005

N2 - Advances in our understanding of biology at the molecular level are very much driven by improvements in the scientist's tool box. Such improvements may not only be an introduction of new techniques like polymerase chain reaction, but as much an increment of for example the sensitivity of existing methods. The in vitro generation of antibodies using phage display is one such technique, which continuously has been developed since its introduction more than 10 yr ago. As a result, selection of phage-displayed antibodies is emerging as a proteomic tool for the identification of differentially expressed cell surface antigens. Here, a method is described that enables the rapid isolation of a panel of recombinant antibodies recognizing epidermal skin keratinocytes. The method exploits the properties of a protease sensitive helper phage and facilitates the isolation of affinity-binders after a single round of selection. This assures a high diversity of binders owing to the reduction of experimental noise.

AB - Advances in our understanding of biology at the molecular level are very much driven by improvements in the scientist's tool box. Such improvements may not only be an introduction of new techniques like polymerase chain reaction, but as much an increment of for example the sensitivity of existing methods. The in vitro generation of antibodies using phage display is one such technique, which continuously has been developed since its introduction more than 10 yr ago. As a result, selection of phage-displayed antibodies is emerging as a proteomic tool for the identification of differentially expressed cell surface antigens. Here, a method is described that enables the rapid isolation of a panel of recombinant antibodies recognizing epidermal skin keratinocytes. The method exploits the properties of a protease sensitive helper phage and facilitates the isolation of affinity-binders after a single round of selection. This assures a high diversity of binders owing to the reduction of experimental noise.

KW - Bacteriophage M13/genetics

KW - Enzyme-Linked Immunosorbent Assay

KW - Epidermis/immunology

KW - Gene Library

KW - Humans

KW - Immunoglobulin Fragments/immunology

KW - Keratinocytes/immunology

M3 - Journal article

C2 - 15502198

VL - 289

SP - 359

EP - 370

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

ER -