Functional analysis of recombinant human serum albumin domains for pharmaceutical applications

Functional analysis of recombinant human serum albumin domains for pharmaceutical applications

Functional analysis of the three recombinant human serum albumin (rHSA) domains and their potential as stand-alone proteins for use as drug delivery protein carriers. Protein structure was examined by fluorescence and CD spectroscopy. Ligand binding was estimated by ultrafiltration. Antioxidant acti...

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Bibliographic Details
Published in:Pharmaceutical research Vol. 21; no. 10; pp. 1924 - 1932
Main Authors: Matsushita, Sadaharu, Isima, Yu, Chuang, Victor Tuan Giam, Watanabe, Hiroshi, Tanase, Sumio, Maruyama, Tomu, Otagiri, Masaki
Format: Journal Article
Language:English
Published: United States Springer Nature B.V 01-10-2004
Subjects:
Algorithms
Animals
Antioxidants
Antioxidants - chemistry
Binding Sites
Buffers
Circular Dichroism
Drug Carriers - chemistry
Drug Carriers - pharmacokinetics
Drug Delivery Systems
Drugs
Esterases - chemistry
Humans
Hydrolysis
Indium Radioisotopes
Ligands
Male
Mice
Peptides
Pharmaceuticals
Pharmacokinetics
Phosphopyruvate Hydratase - chemistry
Pichia - chemistry
Protein Binding
Protein Conformation
Proteins
Reactive Oxygen Species
Recombinant Proteins - chemistry
Recombinant Proteins - pharmacokinetics
Serum Albumin - chemistry
Serum Albumin - pharmacokinetics
Spectrophotometry, Ultraviolet
Tissue Distribution
United States > US
Japan
Malaysia
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Summary:Functional analysis of the three recombinant human serum albumin (rHSA) domains and their potential as stand-alone proteins for use as drug delivery protein carriers. Protein structure was examined by fluorescence and CD spectroscopy. Ligand binding was estimated by ultrafiltration. Antioxidant activity was estimated by measuring the quenching of dihydrorhodamine 123. Esterase-like activity and enolase-like activity were estimated from the rate of hydrolysis of p-nitrophenyl acetate and conversion of dihydrotestosterone from the 3-keto to 3-enol form, respectively. The domains of human serum albumin (HSA) were radiolabeled with 111In to evaluate their pharmacokinetics. The ligand binding ability of subsites Ia and Ib could not be detected in domain II. However, the binding of ligands to subsite Ic and site II were preserved in domain II and domain III, respectively. Domain III retained about 45% of its esterase-like activity, and weaker esterase-like activity was also observed in domain I. All domains showed low enolase-like activity in a pH 7.4 phosphate buffer, but domain II had higher activity in a pH 9.2 carbonate buffer. Domain I exhibited antioxidant activity comparable to that of rHSA. All three of the 111In-radiolabeled domains were rapidly eliminated from HSA, with high accumulation in the kidneys. Domain I of HSA has great potential for further development as a drug delivery protein carrier, due to its favorable properties and the presence of a free cysteine residue.
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content type line 23
ISSN:0724-8741
1573-904X
DOI:10.1023/b:pham.0000045248.03337.0e