Subscriber access provided by UNIV OF WESTERN ONTARIO
Article
Real-time tracking the synthesis and degradation of albumin in complex biological systems with a near-infrared fluorescent probe Qiang Jin, Lei Feng, Shui-Jun Zhang, Dan-Dan Wang, Fangjun Wang, Yi Zhang, Jing-Nan Cui, Wen-Zhi Guo, Guang-Bo Ge, and Ling Yang Anal. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.analchem.7b01975 • Publication Date (Web): 15 Aug 2017 Downloaded from http://pubs.acs.org on August 15, 2017
Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.
Analytical Chemistry is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
Page 1 of 9
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
Analytical Chemistry
Real-time tracking the synthesis and degradation of albumin in complex biological systems with a near-infrared fluorescent probe Qiang Jin†, ‡,⊥, ǁ, §, Lei Feng#,∥,§, Shui-Jun Zhang †, ǁ, Dan-Dan Wang ‡, Fang-Jun Wang⊥, Yi Zhang †, Jing-Nan Cui #, Wen-Zhi Guo*,†, Guang-Bo Ge *,‡,⊥, and Ling Yang ‡ †
Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450001, China ‡
Institute of Interdisciplinary Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, 201213, China ⊥
#
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China
State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian, 116024, China
ǁ
Key Laboratory of Hepatobiliary and Pancreatic Surgery & Digestive Organ Transplantation of Henan Province, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450001, China ∥
College of Pharmacy, Academy of integrative Medicine, Dalian Medical University, Dalian 116044, China *Corresponding Author. E-mail:
[email protected] (G. B. Ge), Tel/Fax: +86-021-51323186;
[email protected] (W.Z. Guo). Tel/Fax: +86-0371-66295358 ABSTRACT: In this study, a novel fluorescent detection system for biological sensing of human albumin (HA) was developed on the basis of the pseudo-esterase activity and substrate preference of HA. The designed near-infrared (NIR) fluorescent probe (DDAP) could be effectively hydrolyzed by HA, accompanied with significant changes in both colour and fluorescence spectrum. The sensing mechanism was fully investigated by fluorescence spectroscopy, NMR and mass spectra. DDAP exhibited excellent selectivity and sensitivity toward HA over a variety of human plasma proteins, hydrolases, and abundant biomolecules found in human body. The probe has been successfully applied to measure native HA in diluted plasma samples and the secreted HA in the hepatocyte culture supernatant. DDAP has also been used for fluorescence imaging of HA reabsorption in living renal cells, and the results show that the probe exhibits good cell permeability, low cytotoxicity and high imaging resolution. Furthermore, DDAP has been successfully used for real-time tracking the uptaking and degradation of albumin in ex vivo mouse kidney models for the first time. All these results clearly demonstrated that DDAP-based assay held great promise for real-time sensing and tracking HA in complex biological systems, which would be very useful for basic researches and clinical diagnosis of HA-associated diseases.
INTRODUCTION Human albumin (HA) is the most abundant plasma protein in the human body with a plasma concentration of 35-55 g/L. HA is synthesized exclusively in hepatocytes as a single polypeptide chain consisting of 585 amino acid residues. Once synthesized, HA is secreted into the plasma as an unmodified form without any glycosylation or other modifications, which has a half-life of 19-21 days in plasma.1 HA participates in regulation of plasma oncotic pressure and transports endogenous and exogenous ligands such as thyroxine, free fatty acids, bilirubin, and various drugs.2-4 In addition to its reversible and irreversible ligand binding capabilities, native HA also exerts several types of hydrolytic activity, among which the esterase-like activity is the most prominent.5-7
determination of HA in biological fluids is of importance in both clinical diagnosis and biomedical researches. In the past 30 years, bromcresol green (BCG) and bromcresol purple (BCP) are widely used for quantification of HA in plasma,12, 13 due to they are simple, sensitive, rapid response, lower cost, non-destruction, high-throughput detection and ease of manipulation. However, such methods lack of specificity and hard to distinguish between native HA and denatured HA. Additionally, such reversible binding may be affected by the high-affinity endogenous interferences (such as bilirubin or bile acids) or exogenous compounds (such as drugs or food chemicals), and thus lead to underestimate or overestimate the levels of HA in real samples.14, 15 In contrast to dye-binding probes, the enzymatic activity-based probes (EABPs) rely on a given enzymatic reaction consisting of recognition, binding and catalysis processes, thus usually display improved selectivity towards target enzyme(s).16-23 Therefore, it is necessary to design and develop a practical activity-based fluorescent probe for highly selective and sensitive quantification of native HA in complex biological systems, such as living cells. Although a few fluorescent EABPs for HA have been developed,24, 25 all of them emit their light at short
In the past half-century, many studies have been clearly shown that the levels of HA in body fluids are closely associated with the progression of many human diseases, including post-menopausal obesity, kidney disease, diabetes mellitus, hypertension and liver injury.8-10 A low HA level in serum, known as hypoproteinemia, may indicate liver failure, cirrhosis, and chronic hepatitis, while an elevated level of HA in urine may indicate kidney diseases.11 Hence, the precise quantitative
1 ACS Paragon Plus Environment
Analytical Chemistry
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
Page 2 of 9
fourier transform ion cyclotron resonance mass spectrometer (LTQ Orbitrap XL). The UV–vis spectra and fluorescence emission/excitation spectra were measured on Synergy H1 Hybrid Multi-Mode Microplate Reader (BioTek). A Shimadzu UFLC system equipped with an SPD-M 20A diode array detector (DAD) and a mass spectrometer was used to determine DDAP and its hydrolytic product. LC-MS/MS based HA quantification was performed on an AB Sciex QTrap® 5500 system with a Turbo VTM source (Framingham, MA, USA).
wavelengths (
