Glycerol diversifies phage repertoire selections and lowers non‐specific phage absorption

FEBS Letters

Kahns, Liselotte¹  Ravn, Peter¹  Wind, Troels¹  Clark, Brian F.C¹  Hobolt Jensen, Kristian¹  Stausbøl-Grøn, Brian¹  Kjær, Svend¹ 

[1] Institute of Molecular and Structural Biology, University of Aarhus, Gustav Wieds Vej 10C, 8000 Århus C, Denmark

关键词: Phage antibody;    Selection strategy;    Glycerol;    Elongation factor 1A;    Dominant clone;    Ag;    antigen;    Amp;    ampicillin;    c.f.u.;    colony forming units;    eEF1A;    eukaryotic elongation factor 1A;    ELISA;    enzyme-linked immunosorbent assay;    Glu;    glucose;    Gly;    glycerol;    HCDR3;    third complementarity determining region of the variable heavy chain;    HRP;    horseradish peroxidase;    LDH;    lactate dehydrogenase;    OPD;    ortho-phenylen diamine;    PBS;    phosphate-buffered saline;    PCR;    polymerase chain reaction;    Phage-ab;    phage displaying antibody fragment;    scFv;    single-chain fragment variable;    VH;    variable region of the heavy chain;    VL;    variable region of the light chain;   

DOI  :  10.1016/S0014-5793(98)00789-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using a semi-synthetic phage displayed antibody repertoire, isoform-specific and cross-reactive phage-antibodies to eukaryotic elongation factor 1A (eEF1A) have been selected. Enrichment of specific antibodies was found to depend on the presence of glycerol. Further selections against lactate dehydrogenase (LDH) revealed that the dominance of a phage-antibody clone to LDH was inhibited by glycerol, a notable feature for selection strategies where a broad variety of binding clones is desired. The impact of glycerol in distinct steps of the selection protocol was examined and glycerol found to affect certain antibody-antigen interactions. Furthermore, the non-specific phage binding was lowered by three orders of magnitude at a 20% (v/v) glycerol concentration.

【 授权许可】

Unknown