Bright oligothiophene N-succinimidyl esters for efficient fluorescent labeling of proteins and oligonucleotides.
The synthesis of multicolor fluorescent oligothiophene N-succinimidyl esters (TSEs) is reported, and their optical properties are discussed with the aid of ab initio calculations. The esters were coupled to proteins and to 3'-amino-modified oligonucleotides in mild conditions and with similar modalities. A comparative study of the bioconjugate of IgG1 anti-CD3 antibody labeled with a blue fluorescent TSE and with fluorescein isothiocyanate (FITC) is reported, showing that the former achieves higher photoluminescence intensity and optical stability than the latter. Fluorescence resonance energy transfer experiments with TSE-labeled oligonucleotides and examples of cellular imaging via TSE-labeled proteins are reported.
Barbarella
Giovanna
G
Consiglio Nazionale Ricerche (ISOF), Mediteknology srl, Area Ricerca CNR, Via Gobetti 101, I-40129 Bologna, Italy. barbarella@isof.cnr.it
Zambianchi
Massimo
M
Ventola
Alfredo
A
Fabiano
Eduardo
E
Della Sala
Fabio
F
Gigli
Giuseppe
G
Anni
Marco
M
Bolognesi
Andrea
A
Polito
Letizia
L
Naldi
Marina
M
Capobianco
Massimo
M
D000911:Antibodies, Monoclonal / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D004705:Endocytosis; D004952:Esters / Q000138:chemical synthesis* / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D000922:Immunotoxins / Q000378:metabolism; D019169:Jurkat Cells; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism; D011485:Protein Binding; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins; D013876:Thiophenes / Q000138:chemical synthesis* / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D017252:CD3 Complex; D004952:Esters; D005456:Fluorescent Dyes; D000922:Immunotoxins; D009841:Oligonucleotides; D010940:Plant Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D013876:Thiophenes; D009699:N-Glycosyl Hydrolases; D000076985:Saporins