2000 Fiscal Year Final Research Report Summary
Design and Evaluation of Albumin Mutants for Clinical Application
| Project/Area Number |
11694298
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biological pharmacy
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| Research Institution | KUMAMOTO UNIVERSITY |
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Principal Investigator |
OTAGIRI Masaki Fac.Pharm.Sci., KUMAMOTO UNIVERSITY Prof., 薬学部, 教授 (80120145)
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| Co-Investigator(Kenkyū-buntansha) |
TANASE Sumio Sch.Med.Sci., KUMAMOTO UNIVERSITY Asso.Prof., 医学部, 助教授 (20112401)
KAI Hirofumi Fac.Pharm.Sci., KUMAMOTO UNIVERSITY Assoc.Prof., 薬学部, 助教授 (30194658)
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| Project Period (FY) |
1999 – 2000
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| Keywords | Albumin preparation / Site-directed mutagenesis / Molecular design / Artificial blood / Functional evaluation / Blood preparation / Clinical application |
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| Research Abstract |
International collaboration work for reducing the rate of albumin preparations usage was undertaken to develop a safe and effective recombinant albumins. The results obtained are as follow :
1) Two single-mutants (K199A, K525A) were produced. The recombinant albumins were correctly folded, as evidenced by the fact that they showed similar patterns as the native albumin in the far- and near-UV CD spectrum as well as reactivity towards polyclonal anti-serum raised against native albumin.
2) Thermal denaturation experiments by using GdnCl and DSC revealed no significant difference in conformational stability for native and mutant albumins.
3) The ligand binding properties and esterase-like activity of the albumin mutants were nearly the same as those for the native albumin.
4) The plasma levels of the albumin mutants labeled with ^<125>I after intravenous bolus injections to rats showed two-phase profiles but their half-lives were shorten than that for the native albumin.
5) Dr.Kragh-Hansen of Aarhus University visited Kumamoto University twice (Oct.23-Nov.7, 1999 ; May 5-Oct.29, 2000) to discuss about pharmacokinetic and biological properties of the albumin mutants. Dr.Curry of Imperial College also visited Kumamoto University (Sep.27-Oct.26, 2000) to discuss about structural characterization of the albumin mutants.
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