Dicyclohexylcarbodiimide → Folate diaminohexane

Beteiligte Stoffe

Reaktionsschritte

8 mg NHS (N-Hydroxysuccinimide) was added to 16 mg folic acid in 4 ml DMSO solvent followed by 9 mg of DCC (dicyclohexyl carbodiimide) in 500 μl DMSO. It was then incubated at room temperature for 2 h then added to 4.7 mg Diaminohexane in 500 μl DMSO forming a white-yellow precipitate/suspension. It was left overnight to react to completion forming the folate-diaminohexane linker. 156 μl of this was then added to 1 mg of Sulfo-SBED (2-[(6-(Biotinamido)-2-(p-azidobenzamido)-hexanamido]ethyl-1-3′-dithio-propionate) (Pierce & Warriner product number 33033) in 100 μl DMF and incubated for a further 48 h during which time the solution clarified. This was followed by the addition of 750 μl distilled water to neutralise any tridentate-NHS ester groups which might have remained. After a further 4 h 500 μl of this hydrolysed tridentate-folate complex was added to 1 mg (500 μl) of antibody and incubated for a further 48 h.

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