I need to develop a paper based assay to determine urinary creatinine levels. Since creatinine is a small molecule, I need to conjugate it to a protein like BSA to make it suitable for the assay.
Has anyone successfully able to conjugate creatinine to BSA ?
Thanking you.
Hi Gangula,
You might try activating carboxyl groups in BSA with NHS (N-hydroxysuccinimide) or sulfo-NHS reagent (similar protocol could be used as for amine-coupling in e.g. SPR), which then should selectively form an amide bound with the NH2 functionality of creatinine. Add "activated" BSA in excess to react all creatinine for quantitation. After a few minutes (should be optimized), block unreacted BSA and NHS with excess 1M ethanolamine. I attach an example reaction scheme.
Hope this helps,
Karoly
- Badges
- Science topic