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Isolation and characterization of single‐chain Fv genes encoding antibodies specific for Drosophila Poxn protein
Author(s) -
Hassanzadeh Gh Gholamreza,
De Silva Kumudu S.K.,
Dambly-Chaudière Christine,
Brys Lea,
Ghysen Alain,
Hamers Raymond,
Muyldermans Serge,
De Baetselier Patrick
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)01204-6
Subject(s) - recombinant dna , microbiology and biotechnology , gene , antibody , function (biology) , schneider 2 cells , biology , alpha (finance) , chemistry , biochemistry , genetics , rna interference , rna , medicine , construct validity , nursing , patient satisfaction
The usefulness of intrabodies as specific inhibitors of gene function has been extensively demonstrated in cell culture assays. However, very few experiments have been conducted with intrabodies expressed in whole organisms. To evaluate the intrabody technology in Drosophila , we focused on poxn protein, since its effects can be easily studied. We purified the recombinant poxn protein. We next isolated three single‐chain variable fragments (scFv) which specifically recognize poxn protein. Two scFvs, designated α‐Poxn2 and α‐Poxn4, react with both denatured and native Poxn with half maximal inhibition values of 100 nM and 40 nM, respectively. The α‐Poxn5 scFv also recognizes denatured Poxn but either does not recognize native Poxn or its half maximal inhibition value for native Poxn is high.