A model phage display subtraction method with potential for analysis of differential gene expression

FEBS Letters

Kahns, Liselotte¹  Clark, Brian F.C.¹  Wind, Troels¹  Stausbøl-Grøn, Brian¹  Kjær, Svend¹  Hansen, Nils J.V.¹  Kristensen, Peter¹ 

[1] Institute of Molecular and Structural Biology, Biostructural Chemistry, University of Aarhus, Langelandsgade 140, 8000 Aarhus C, Denmark

关键词: Phage display;    Phage display subtraction;    Competitive selection;    Phage antibody library;    Antibody engineering;    Single-chain Fv fragment;    A 490mm;    UV absorption at 490 nm;    cfu;    colony-forming units;    EDTA;    ethylenedinitrilotetraacetic acid;    ELISA;    enzyme-linked immunosorbent assay;    IPTG;    isopropyl-l-thio-β-d-galactopyranoside;    LDH;    lactate dehydrogenase;    MES;    2-[N-morpholino]ethanesulphonic acid;    MOPS;    3-[N-morpholino]propanesulphonic acid;    MPBS;    skimmed milk powder in PBS;    PBS;    phosphate-buffered saline;    PMSF;    phenylmethylsulphonyl fluoride;    scFv;    single-chain Fv fragment;   

DOI  :  10.1016/0014-5793(96)00703-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In order to establish a subtractive procedure that makes it possible to enrich selectively phage displayed antibodies directed against proteins constituting a difference between two populations of cells, a competitive selection strategy utilising two solid phases was developed and tested. Antibodies recognising a defined difference between two otherwise identical protein mixtures were isolated and their specificity confirmed. To test further the efficacy of selection inhibition during the competitive selections, selections towards a total cell extract were performed with and without competition from the same extract. An analysis of the resulting phage antibodies confirmed the subtractive nature of the system described.

【 授权许可】

Unknown