The phototrityl group. Photocrosslinking of oligonucleotides to BSA

Bioorganic and Medicinal Chemistry Letters

Volumn 6, Issue 10, 1996, Pages 1167-1170

  Bidaine, A ^a   Berens, C ^a   Sonveaux, E ^a  

^a UNIVERSITÉ CATHOLIQUE DE LOUVAIN   (Belgium)

Author keywords

[No Author keywords available]

Indexed keywords

ANTISENSE OLIGONUCLEOTIDE; BOVINE SERUM ALBUMIN; DNA; OLIGONUCLEOTIDE; TRITYL DERIVATIVE;

ADSORPTION; ARTICLE; CONJUGATION; CROSS LINKING; HYBRIDIZATION; IMMOBILIZATION; TARGET ORGAN;

BOVINAE;

EID: 0030060917     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/0960-894X(96)00191-6     Document Type: Article

References (21)

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14. 18 reversed phase HPLC (see note 15). It was homogeneous in PAGE.
15. 3CN 60%. Gradient from 5% B to 60% B in 1h. Flow rate 1 ml/min.). Oligonucleotide 7 featured a retention time of 26 min. and 8, a retention time of 42 min. Several peaks coming from the hydrolysis of the acylating agent were also present. They could be removed by ethanol precipitation of the oligonucleotide at -80°c (0.5 h). The acylated product 8 was isolated by prep-HPLC. It had the correct size in PAGE.
16. Connors, K. A. In Remington: The Science and Practice of Pharmacy; Gennaro, A. R., Ed.; Mack Publishing Company: Easton, Pennsylvania, 1995; Vol. 1, p. 181.
17. Bovine albumin was Cohn V fractionate (ethanol precipitated) from Sigma. The monomeric fraction was isolated by gel chromatography on a G-200 column of 2.5 cm width and 1 m length. The eluent was PBS. The oligonucleotide was incubated with BSA in PBS at 20°C for two hours. A complementary experiment, with compound 7, where X = 2-nitrobenzoyl instead of H, showed that trityl capped oligonucleotides were fully adsorbable on BSA. The percentage of oligonucleotide adsorbed (7, with X = 2-nitrobenzoyl, 1 μg of oligomer per 100 μl of PBS) was 61, 73, 89, 91, 92 and 96% at molar ratios oligonucleotide:BSA, 1:0.5, 1:1, 1:2, 1:5, 1:20 and 1:50, respectively.
18. 3CN 10%. Eluent B = same, but 2M NaCl. Gradient from 0% B to 100% B in 1h. Flow rate 0.5 ml/min. BSA had a retention time of 9 min and the conjugate had a retention time of 13 min.
19. The ExcelGel™ (gradient 8-18) precasted gel from Pharmacia was used as recommended by the supplier.
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