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Marani, M.M.; Ceron, M.C.M.; Giudicessi, S.L.; De Oliveira, E.; Côté, S.; Erra-Balsells, R.; Albericio, F.; Cascone, O.; Camperi, S.A. "Screening of one-bead-one-peptide combinatorial library using red fluorescent dyes. Presence of positive and false positive beads" (2009) Journal of Combinatorial Chemistry. 11(1):146-150
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Abstract:

To screen one-bead-one-compound (OBOC) combinatorial libraries, tens of thousands to millions of compound beads are first mixed with a target molecule. The beads that interact with this molecule are then identified and isolated for compound structure determination. Here we describe an OBOC peptide library screening using streptavidin (SA) as probe protein, labeled with a red fluorescent dye and using the COPAS BIO-BEAD flow sorting equipment to separate fluorescent from nonfluorescent beads. The red dyes used were ATTO 590 and Texas Red. After incubating the library with the SA-red fluorescent dye conjugate, we isolated positive beads caused by peptide-SA interaction and false positive beads produced by peptide-fluorescent dye interaction. These false positives were a drawback when sorting beads by COPAS. However, an in depth analysis of both kinds of beads allowed the differentiation of positives from false positives. The false positive beads showed bright homogeneous fluorescence, while positive beads had a heterogeneous fluorescence, exhibiting a characteristic halo appearance, with fluorescence intensity greatest at the bead surface and lowest in the core. The difference was more evident when using Texas Red instead of ATTO 590. Thus, positive beads could be manually separated from false positive ones. The beads were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Most of the sequences obtained from positive beads had the His-Pro-Gln motif. Peptides from false positive beads were rich in Leu/Ileu, His, Phe, and Tyr. © 2009 American Chemical Society.

Registro:

Documento: Artículo
Título:Screening of one-bead-one-peptide combinatorial library using red fluorescent dyes. Presence of positive and false positive beads
Autor:Marani, M.M.; Ceron, M.C.M.; Giudicessi, S.L.; De Oliveira, E.; Côté, S.; Erra-Balsells, R.; Albericio, F.; Cascone, O.; Camperi, S.A.
Filiación:Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, (1113) Ciudad Autónoma de Buenos Aires, Argentina
CIHDECAR-CONICET, Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pabellón II-Ciudad Universitaria, (1428) Ciudad Autónoma de Buenos Aires, Argentina
Proteomics Platform, Barcelona Science Park, Baldiri Reixac 10, 08028 Barcelona, Spain
Matrix Innovation Inc., 945 Newton, Québec City, QC, Canada
Institute for Research in Biomedicine, Barcelona Science Park, Baldiri Reixac 10, 08028 Barcelona, Spain
CIBER-BBN, Networking Centre on Bioengineering, Biomaterials and Nanomedicine, Barcelona Science Park, Baldiri Reixac 10, 08028 Barcelona, Spain
Palabras clave:fluorescent dye; microsphere; peptide library; article; combinatorial chemistry; drug screening; methodology; Combinatorial Chemistry Techniques; Drug Evaluation, Preclinical; Fluorescent Dyes; Microspheres; Peptide Library
Año:2009
Volumen:11
Número:1
Página de inicio:146
Página de fin:150
DOI: http://dx.doi.org/10.1021/cc800145c
Título revista:Journal of Combinatorial Chemistry
Título revista abreviado:J. Comb. Chem.
ISSN:15204766
CODEN:JCCHF
CAS:Fluorescent Dyes; Peptide Library
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15204766_v11_n1_p146_Marani

Referencias:

  • The following abbreviations are used: ACN, acetonitrile; AcOH, acetic acid; BSA, bovine serum albumin; BSA-TR, bovine serum albumin-Texas Red; CHCA, α-cyano-4-hydroxycinnamic acid; DCM, dichloromethane; DIPCDI, N, N-1,3-diisopropylcarbodiimide; DIPEA, N,N-diisopropylethylamine; DMAP, 4-(N,N-dimefhylamino)pyridine; DMF, N,N-dimethylformamide; FITC, fluorescein isothiocyanate; Fmoc-amino acids, fluorenylmethoxyearbonylamino acids; HMBA, 4-hydroxymethylbenzoic acid; HOBt, 1-hydroxybenzotriazole; MALDI-TOF MS, matrix-assisted laser desorption ionization time-of-flight mass spectrometry; OBOC, one-bead one-compound; PBS, phosphate-buffered saline; PEG, poly(ethyleneglycol, SA, streptavidin; SA-ATTO, streptavidin ATTO 590 conjugate; SA-POD, streptavidin peroxidase conjugate; SA-TR, streptavidin Texas red conjugate; TBS, Tris buffer saline; TBTU, 1-[bis(dimethylamino)methylene]-1H-benzotriazolium hexafluorophosphate N-oxide; TFA, triflouroaceti; Lam, K.S., Salmon, S.E., Hersh, E.M., Hruby, V.T., Kazmierski, W.M., Knapp, R.J., (1991) Nature, 354, pp. 82-84
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Citas:

---------- APA ----------
Marani, M.M., Ceron, M.C.M., Giudicessi, S.L., De Oliveira, E., Côté, S., Erra-Balsells, R., Albericio, F.,..., Camperi, S.A. (2009) . Screening of one-bead-one-peptide combinatorial library using red fluorescent dyes. Presence of positive and false positive beads. Journal of Combinatorial Chemistry, 11(1), 146-150.
http://dx.doi.org/10.1021/cc800145c
---------- CHICAGO ----------
Marani, M.M., Ceron, M.C.M., Giudicessi, S.L., De Oliveira, E., Côté, S., Erra-Balsells, R., et al. "Screening of one-bead-one-peptide combinatorial library using red fluorescent dyes. Presence of positive and false positive beads" . Journal of Combinatorial Chemistry 11, no. 1 (2009) : 146-150.
http://dx.doi.org/10.1021/cc800145c
---------- MLA ----------
Marani, M.M., Ceron, M.C.M., Giudicessi, S.L., De Oliveira, E., Côté, S., Erra-Balsells, R., et al. "Screening of one-bead-one-peptide combinatorial library using red fluorescent dyes. Presence of positive and false positive beads" . Journal of Combinatorial Chemistry, vol. 11, no. 1, 2009, pp. 146-150.
http://dx.doi.org/10.1021/cc800145c
---------- VANCOUVER ----------
Marani, M.M., Ceron, M.C.M., Giudicessi, S.L., De Oliveira, E., Côté, S., Erra-Balsells, R., et al. Screening of one-bead-one-peptide combinatorial library using red fluorescent dyes. Presence of positive and false positive beads. J. Comb. Chem. 2009;11(1):146-150.
http://dx.doi.org/10.1021/cc800145c