Development And Biological Application Of Human Serum Albumin Fluorescence Probe

Background: Human serum albumin(HSA)is the most abundant protein in human plasma.Albumin accounts for 50-60 percent of total serum protein in plasma.HSA plays key roles in human health.HSA is produced exclusively by hepatic cells.It is secreted into the portal circulation as soon as it is synthesized.The concentration of HAS is 35-55 g/L in plasma of healthy adult people.Moreover,many diseases may lead to the decline of albumin level.HSA is commonly measured clinically as a part of blood routine test as an important diagnostic and prognostic marker for many related diseases.Therefore,the correct determination of HSA is of significant importance in clinical application.Aim: To develop a fluorescence probe ACDM for simple,accurate,sensitive,anti-interferential,good biocompatibility and low cost detection of HSA and to evaluate its biological assessment and the application on the detection of the biological samples.Methods:(1)Confirmation of ACDM chemical and physical stability(including p H,organic solvent,temperature,binding rate etc);(2)The protein selectivity and anti-interference upon endogenous and exogenous substances of ACDM;(3)Assignment of binding site(s)of ACDM on HSA with the jobs plot method and drug site markers;using the Molecular docking and molecular dynamics simulations to make the confirmation;(4)HSA detection in 28 plasma and Hep G2 cell culture supernatant by ACDM assays;using the BCG and proteomics method to make the comparison.Results:(1)The optimal p H is 8.0.The volume of DMSO is 1%.The final ACDM concentration is 20 μM.Keeping the temp in room temperature and reacting 3 s;(2)the formation of ACDM-HA complex leads to the occurrence of a strong fluorescence response and has good anti-interference;(3)ACDM could bind on HA steadily via binding on a non-drug binding site.(4)ACDM could be used to determine HSA in plasma and Hep G2 cell culture supernatant.Moreover the quantification was highly inconsistent with the control method.Conclusions: A rapid-response fluorescent probe ACDM for highly selective,sensitive,anti-interferential detection of human albumin in complex biological samples has been developed.Moreover the quantification was quite highly reliable.Therefore,ACDM could be used for clinical albumin test for diagnosis and prognosis of related disease.